Saframycins, analogues and uses thereof

ABSTRACT

In recognition of the need to develop novel therapeutic agents and efficient methods for the synthesis thereof, the present invention provides novel compounds of general formula (I), and methods for the synthesis thereof.  
                 
In another aspect, the present invention provides pharmaceutical compositions comprising a compound of formula (I) and a pharmaceutically acceptable carrier. In yet another aspect, the present invention provides methods for treating cancer comprising administering a therapeutically effective amount of a compound of formula (I) to a subject in need thereof.

PRIORITY INFORMATION

The present application claims priority under 35 U.S.C. §119(e) toprovisional application number 60/245,888, filed November 3, 2000,entitled “Synthesis of Saframycins, Analogues and Uses Thereof”, theentire contents of which are hereby incorporated by reference.

GOVERNMENT SUPPORT

This invention was made in part with a grant from the NationalInstitutes of Health (Grant Number: 7 R37 CA47148-12). Therefore, thegovernment has certain rights in the invention.

BACKGROUND OF THE INVENTION

The discovery of novel therapeutic agents has traditionally relied onthe identification of biologically active secondary metabolites ofmicroorganisms. These compounds have provided a rich source of naturalproducts that have either been utilized directly as effectivetherapeutic agents, or have provided leads for novel therapeutic agentsto be developed through synthetic techniques.

One disease for which the development of novel therapeutics isparticularly important is cancer, which not only has eluded a “cure”,but is also one of the leading disease-related causes of death of thehuman population. Examples of anticancer agents that have beenidentified from or developed from natural sources include paclitaxel,mitomycin C, and adriamycin to name a few. One drawback to the use ofsecondary metabolites from natural resources, however, has been thatthese agents are generally only present in minute quantities.Fortunately, in an effort to make these agents more available for use,and to enable further pharmaceutical research, synthetic chemists havedeveloped elegant and efficient synthetic strategies to enable theproduction of either the natural products themselves, or usefulderivatives thereof.

Although these therapeutic agents, and others developed from naturalsources, through the efforts of synthetic chemistry, are currently inuse for the treatment of individuals having cancer, many of theseagents, as well as other common treatments such as surgery andradiation, are often unselective for tumor cells and/or are so toxic asto render the individual significantly immunocompromised. Thus, althoughmany strides have been made in the development of novel treatments,there remains a need for the identification of additional therapeutics,preferably those that are more selective and less toxic.

One particular family of natural products that has generated significantinterest is the saframycins. The saframycins are a class of antibioticswith activity against gram-positive bacteria and also against severalkinds of tumor. Specifically, several saframycins analogues have beenisolated and characterized in recent years (see, DE 2839668; U.S. Pat.Nos. 4,248,863; 4,372,947; 5,023,184; 4,837,149; and EP 329606). Forexample saframycins A-H, R and S have been isolated from the culturebroths of Streptomyces lavendulae, and saframycins M_(x1) and M_(x2),have been isolated from the culture broths of the myxobacterium,Myxococcus xanthus, each of the saframycins varying in the oxidationstate of the ring system and in substitution of the core structure (see,for example, Saito et al. Chem. Pharm. Bull. 1995, 43, 777). It has beensuggested that certain saframycins, namely A and C exhibit extremecytotoxicity toward culture cells and toward several experimental tumorsincluding leukemias L1210 and P388 and Ehrlich carcinoma. Specifically,saframycin A has been shown to block RNA synthesis in cultured cells,and it has been suggested that saframycins A and C exhibit this potencybecause of their ability to bind and cleave DNA (for a discussion of thebiological activity of saframycins see, for example, Lown et al.Biochemistry 1982, 21, 419; Ishiguro et al. Biochemistry 1978, 17, 2545;Rao et al. Chem. Res. Toxicol 1990, 3, 262; Ishiguro et al. J Biol.Chem. 1981, 256, 2162). Although this class of natural products hasshown promising biological activity, there have been few investigationsinto the synthesis and development of novel analogues of this family ofnatural products (see, EP 233841; EP 173649; Fukuyama et al. J. Am.Chem. Soc. 1982, 104, 4957-4958; Kubo et al. J. Org. Chem. 1988, 53,4295-4310; Fukuyama et al. J. Am. Chem. Soc. 1990,112, 3712-3713).

Clearly, there remains a need to further investigate the potential ofthis class of natural products, and analogues thereof, to develop novel,more potent and more selective therapeutics. Additionally, because ofthe complexity of the structure of this class of natural products, therealso remains a need to develop additional synthetic techniques torapidly access novel compounds based upon the general core structure ofthe saframycins, and other related compounds.

SUMMARY OF THE INVENTION

In recognition of the need to develop novel therapeutic agents andefficient methods for the synthesis thereof, the present inventionprovides novel compounds of general formula (I), and methods for thesynthesis thereof.

The present invention additionally provides pharmaceutical compositionscomprising a compound of formula (I) and a pharmaceutically acceptablecarrier. In yet another aspect, the present invention provides methodsfor treating cancer comprising administering a therapeutically effectiveamount of a compound of formula (I) to a subject in need thereof.

DESCRIPTION OF THE FIGURES

FIG. 1 depicts a retrosynthetic analysis of saframycin A.

FIG. 2 depicts the synthesis of saframycin A and precursors. Reactionconditions: (a) Na₂SO₄, CH₂Cl₂, 23° C., >90%; LiBr, DME, 35 C, 65-72%.(b) CH₂O—H₂O, NaBH(OAc)₃, CH₃CN, 23° C, 94%. (c) HOAc, TBAF, THF, 23°C.; DBU, CH₂Cl₂, 23° C., 92%. (d) Na₂SO₄, CH₂Cl₂, 23° C, 66%. (e) ZnCl₂,TMSCN, CF₃CH₂OH-THF, 23° C., 86%. (f) DBU, CH₂Cl₂, 23° C. 88%. (g)ClCOCOCH₃, PhNEt₂, CH₂Cl₂, 0° C., 89%. (h) PhIO, CH₃CN—H₂O, 0° C., 66%.

FIG. 3 depicts the synthesis of a precursor of safiamycin A (9) from anN-linked trimeric α-amino aldehyde precursor.

FIG. 4 depicts precursors in the synthesis of saframycin A.

FIG. 5 depicts the synthesis of an N-linked trimeric α-amino aldehydeprecursor (8).

FIG. 6 depicts the synthesis of a precursor of saframycin A (9) from anN-linked trimeric α-amino aldehyde precursor (8).

FIG. 7 depicts the synthesis of exemplary inventive analogues ofsaframycin.

FIG. 8 depicts depicts the synthesis of exemplary inventive analogues ofsaframycin.

FIG. 9 depicts depicts the synthesis of exemplary inventive analogues ofsaframycin.

FIG. 10 depicts a general method for the rapid synthesis of largenumbers of potent derivatives of saframycin A.

FIG. 11 depicts the solid phase synthesis of derivatives of saframycinA. Reaction conditions: (a) Imidazole, DMF, 23° C; CH₃OH, Imidazole, 23°C, 100% (2 steps). (b) TBAF, AcOH, THF, 23° C. (c) Piperidine, DMF, 23°C. (d) aldehyde 1, DMF, 23° C. (e) LiBr, DME, 35° C., 83% (4 steps). (f)CH₂O—H₂O, NaBH(OAc)₃, DMF, 23° C., 95%. (g) Fmoc-glycinal, DCE, 40° C.,80% (3 steps). (h) ZnCl₂, TMSCN, CF₃CH₂OH-THF, 23° C, 48%. (*=4.5:1mixture of diastereomers (cis- is major))

FIG. 12 depicts the synthesis of the siloxymorpholine reagent (5).Reaction conditions: (a) Jacobsen HKR (Tokunaga, M.; Larrow, J. F.;Kakiuchi, F.; Jacobsen, E. N. Science. 1997, 277, 936-938). 23° C., 100%(98% ee). (b) 2-aminoethanol, EtOH, 0→70° C, 100%. (c) BnBr, KHCO₃, DMF,50° C., 97%. (d) NaH, THF, 0→23° C.; TsIm, THF, 0→23° C., 70% (2steps, >95% ee). (e) 10% Pd/C, H₂, CH3OH, AcOH, 23° C., 96%.

FIG. 13 depicts reaction with a substituted aldehyde reagent (viaPictet-Spengler reaction) to generate diversified pentacyclic corestructures.

FIGS. 14A and 14B depict a synthetic scheme illustrating the generationof diversity in the R₃ position of generic structure (1) (via alternateN-alkylation reaction).

DETAILED DESCRIPTION OF THE INVENTION

In recognition of the need to develop novel anticancer therapeutics andmore efficient processes for the preparation of such therapeutics, thepresent invention provides novel compounds and methods for thepreparation thereof. In general, in one aspect, the present inventionprovides novel analogues of the saframycin antitumor antibiotics havinganticancer activity. In yet another aspect, the present inventionprovides efficient methods for the generation of these compounds, andalkaloids in general, involving the directed condensation of substitutedaldehyde precursors. Significantly, the methodology provided by thepresent invention enables the efficient production of these novelcompounds in significant quantities for therapeutic use.

1) General Description of Compounds of the Invention

As mentioned above, in one aspect of the invention, novel analogues ofthe saframycin antitumor antibiotics are provided. In general, compoundshaving the structure (I) are provided:

wherein R₁ is NR_(A)R_(B), —OR_(A), —SR_(A), —C(═O)R_(A—, —C(═)R) _(A),—S(O)₂R_(A), or an aliphatic, heteroaliphatic, aryl, heteroaryl,(aliphatic)aryl, (aliphatic)heteroaryl, (heteroaliphatic)aryl, or(heteroaliphatic)heteroaryl moiety, wherein each occurrence of R_(A) andR_(B) is independently hydrogen, —(C═O)R_(C), —NHR_(C), —(SO₂)R_(C),—OR_(C), or an aliphatic, heteroaliphatic, aryl, or heteroaryl moiety,or R_(A) and R_(B), when taken together form an aryl, heteroaryl,cycloaliphatic, or cycloheteroaliphatic moiety, wherein each occurrenceof R_(C) is independently hydrogen, —OR_(D), —SR_(D), —NHR_(D),—(C═O)R_(D), or an aliphatic, heteroaliphatic, aryl, or heteroarylmoiety, wherein each occurrence of R_(D) is independently hydrogen, aprotecting group, or an aliphatic, heteroaliphatic, aryl, heteroaryl,acyl, alkoxy, aryloxy, alkylthio, arylthio, heteroaryloxy, orheteroarylthio moiety;

wherein R₂ is hydrogen, —OR_(E), ═O, —C(═O)RE, —CO₂R_(E), —CN, —SCN,halogen, —SR_(E), —SOR_(E), —SO₂R_(E), —NO₂, —N(RE)₂, —NHC(O)R_(E), oran aliphatic, heteroaliphatic, aryl, or heteroaryl moiety, wherein eachoccurrence of R_(E) is independently hydrogen, a protecting group, or analiphatic, heteroaliphatic, aryl, heteroaryl, acyl, alkoxy, aryloxy,alkylthio, arylthio, heteroaryloxy, or heteroarylthio moiety;

wherein R₃ is hydrogen, a nitrogen protecting group, —COOR_(F),—COR_(F), —CN, or an aliphatic, heteroaliphatic, aryl, or heteroarylmoiety, wherein each occurrence of R_(F) is independently hydrogen, aprotecting group, or an aliphatic, heteroaliphatic, aryl, heteroaryl,alkoxy, aryloxy, alkylthio, arylthio, heteroaryloxy, or heteroarylthiomoiety;

wherein R₄ and R₆ are each independently hydrogen, or an aliphatic,heteroaliphatic, aryl, heteroaryl, acyl, alkoxy, aryloxy, alkylthio,arylthio, heteroaryloxy, or heteroarylthio moiety;

wherein R₅ and R₇ are each independently hydrogen, —ORG, —C(═O)R_(G),—CO₂R_(G), —CN, —SCN, halogen, —SR_(G), —SOR_(G), —SO₂R_(G), —NO₂,—N(R_(G))₂, —NHC(O)R_(G), or an aliphatic, heteroaliphatic, aryl orheteroaryl moiety, wherein each occurrence of R_(G) is independentlyhydrogen, a protecting group, or an aliphatic, heteroaliphatic, aryl,heteroaryl, acyl, alkoxy, aryloxy, alkylthio, arylthio, heteroaryloxy,or heteroarylthio moiety;

wherein R₈ is hydrogen, alkyl, —OH, protected hydroxyl, ═O, —CN, —SCN,halogen, —SH, protected thio, alkoxy, thioalkyl, amino, protected amino,or alkylamino;

wherein m is 0-5;

wherein X₁, X₂, X₃ and X₄ are each independently hydrogen, —OR_(H), ═O,—C(═O)R_(H), —CO₂R_(H), —CN, —SCN, halogen, —SR_(H), —SOR_(H), —SO₂RH,—NO₂, —N(RH)₂, —NHC(O)R_(H), or an aliphatic, heteroaliphatic, aryl, orheteroaryl moiety, wherein each occurrence of R_(H) is independentlyhydrogen, a protecting group, or an aliphatic, heteroaliphatic, aryl,heteroaryl, acyl, alkoxy, aryloxy, alkylthio, arylthio, heteroaryloxy,or heteroarylthio moiety;

whereby if at least either X₁ and X₂ or X₃ and X₄ are doubly bonded tothe 6-membered ring, then the dotted bonds in either or both of the6-membered rings represent two single bonds and one double bond, and aquinone moiety is generated, or if at least either X₁ and X₂ or X₃ andX₄ are singly bonded to the 6-membered ring, then the dotted bonds ineither or both of the 6-membered rings represent two double bonds andone single bond, and a hydroquinone moiety is generated;

whereby each of the foregoing aliphatic, heteroaliphatic and alkylmoieties may independently be substituted or unsubstituted, branched orunbranched, or cyclic or acyclic, and each of the foregoing aryl orheteroaryl moieties may independently be substituted or unsubstituted;and

pharmaceutically acceptable derivatives thereof.

In certain embodiments of the invention, compounds of formula (I) havethe following stereochemistry and structure as shown in formula (Ia):

It will be appreciated that, in certain embodiments of the compounds asdescribed generally above and in classes and subclasses herein certainnaturally occurring saframycins and other related natural products areexcluded including:

-   -   saframycins A, B, C, D, E, F, G, H, R, S₁, Y₃, Y_(d1), A_(d1),        Y_(d2), Y_(2b), Y_(2b-d), AH₂, AH₂Ac, AH₁, AH₁Ac, AR₃, M_(x-1)        and M_(x-2); safracins A and B; reineramycins A, B, D, E, and F;        and xestomycin.

In certain other embodiments, for compounds as described above and insubclasses herein, when m is 1, R₁ excludes any one or more of thefollowing groups: —NH(protecting group), —NH₂, —NHCOCOMe,—NHCOC(Me)(OMe)(OMe), —NHCOCH(NH₂)CH₃, —NHCOCH(NH(acyl))CH₃,—NHCOCHAHH₂)Ac, or NHCOCH(NHCOOBn)(Me); —O(C═O)C(CH₃)═C(CH₃)H; —OH,—O(protecting group), —O(COCH₃), —O(C═O)CH₂CH₃.

In still other embodiments, for certain of the compounds as describedabove and herein, when m is 1; when X₁, X₂, X₃ and X₄ are each ═O; whenR₂ is —CN or —OH; when Hr and are each —CH₃; when R₅ and R₇ are each—OCH₃; when R₈ is H; and R₁ is —NH(C═O)R_(C), then R_(C) is not—CH(NR_(W)R_(Y))(CH₂R_(Z)) where R_(W) and R_(Y) are each independentlyhydrogen or C₁₋₇ alkyl, aryl(C₁₋₄)alkyl, (C₁₋₄)alkylaryl, a substitutedsulfonyl (—S(O)₂—) group, or a substituted acyl group, and where R_(Z)is hydrogen or C₁₋₄ alkyl.

In yet other embodiments, for certain of the compounds as describedabove and herein, when m is 1; when X₁, X₂, X₃ and X₄ are each ═O; whenR₂ is —CN; when R₄ and R₆ are each —CH₃; when R₅ and R₇ are each —OCH₃;when R₈ is H; and R₁ is —NH(C═O)R_(C), then R_(C) is not—C(OH)(Me)CH₂(C═O)Me.

In still other embodiments, for certain of the compounds as describedabove and herein, when m is 1 and when R₂ is H; and R₁ is —NH(C═O)R_(C),then R_(C) is not —CH(Me)NH(C═O)O(CH₂)Ph.

In yet other embodiments, for certain of the compounds as describedabove and herein, when m is 0; R₂ is H; X₃ is H; and R₁ is —C(═O)R_(A),then R_(A) is not —O(alkyl). Alternatively, in certain otherembodiments, when R₂ is H; m is 1; and R₁ is —OR_(A), then R_(A) is not—C(═O)R_(C), or S(O)₂R_(C), wherein R_(C) is an alkyl moiety.

2) Featured Classes of Compounds

It will be appreciated that for compounds as generally described above,certain classes of compounds are of special interest. For example, oneclass of compounds of special interest includes those compounds of theinvention as described above and in certain subclasses herein, whereinthe compounds have the general structure (II):

wherein R₁—R₈, X₁—X₄ and m are as defined above and in subclassesherein.

Another class of compounds of special interest includes those compoundsof the invention as described above and in certain subclasses herein,wherein the compounds have the general structure (III):

wherein R₂—P₈, X₁—X₄, m, R_(A) and R_(B) are as defined above and insubclasses herein.

Yet another class of compounds of special interest includes thosecompounds of the invention as described above and in certain subclassesherein, wherein the compounds have the general structure (IV):

wherein R₂—R₈, X₁—X₄, m, R_(A) and R_(B) are as defined above and insubclasses herein.

Yet another class of compounds of special interest includes thosecompounds of the invention as described above and in certain subclassesherein, wherein the compounds have the general structure (V):

wherein R₂—R₈, X₁—X₄, m, and R_(A) are as defined above and insubclasses herein.

Yet another class of compounds of special interest includes thosecompounds of the invention as described above and in certain subclassesherein, wherein the compounds have the general structure (VI):

wherein R₂—R₈, X₁—X₄, m, and R_(A) are as defined above and insubclasses herein.

Yet another class of compounds of special interest includes thosecompounds of the invention as described above and in certain subclassesherein, wherein the compounds have the general structure (VII):

wherein R₂—R₈, X₁—X₄, m, and R_(A) are as defined above and insubclasses herein.

Yet another class of compounds of special interest includes thosecompounds of the invention as described above and in certain subclassesherein, wherein the compounds have the general structure (VIII):

wherein R₂—R₈, X₁—X₄, m, and R_(A) are as defined above and insubclasses herein.

Still another class of compounds of special interest includes thosecompounds of the invention as described above and in certain subclassesherein, wherein the compounds have the general structure (IX):

wherein R₂—R₈, X₁—X₄, and m are as defined above and in subclassesherein, and wherein R₁ is a substituted or unsubstituted, cyclic oracyclic, branched or unbranched aliphatic or heteroaliphatic moiety, oris a substituted or unsubstituted aryl or heteroaryl moiety.

Yet another class of compounds of special interest includes thosecompounds of the invention as described above and in certain subclassesherein, wherein the compounds have the general structure (X):

wherein R₂—R₈, X₁—X₄, and m are as defined above and in subclassesherein, and wherein R₁ is a substituted or unsubstituted, cyclic oracyclic, branched or unbranched aliphatic or heteroaliphatic moiety, oris a substituted or unsubstituted aryl or heteroaryl moiety.

The following compounds are illustrative of certain of the compoundsdescribed generally and in classes and subclasses herein:

A number of important subclasses of each of the foregoing classesdeserve separate mention; these subclasses include subclasses of each ofthe foregoing classes in which:

i) compounds of the invention as described above and herein areenantiopure;

ii) compounds as described above and in subclasses herein, wherein whenm is 1, R₁ excludes any one or more of the following groups:—NH(protecting group), —NH₂, —NHCOCOMe, —NHCOC(Me)(OMe)(OMe),—NHCOCH(NH₂)CH₃, —NHCOCH(NH(acyl))CH₃ —NHCOCH(NH₂)Ac, orNHCOCH(NHCOOBn)(Me); —O(C═O)C(CH₃)═C(CH₃)H; —OH, —O(protecting group),—O(COCH₃), —O(C═O)CH₂CH₃;

iii) compounds as described above and in subclasses herein, wherein whenm is 1; when X₁, X₂, X₃ and X₄ are each ═O; when R₂ is —CN or —OH; whenR₄ and R₆ are each —CH₃; when R₅ and R₇ are each —OCH₃; when R₈ is H;and R₁ is —NH(C═O)R_(C), then R_(C) is not —CH(NR_(W)R_(Y))(CH₂R_(Z))where R_(W) and R_(Y) are each independently hydrogen or C₁₋₇ alkyl,aryl(C₁₋₄alkyl, (C₁₋₄)alkylaryl, a substituted sulfonyl (—S(O)₂—) group,or a substituted acyl group, and where R_(Z) is hydrogen or C₁₋₄ alkyl;

iv) compounds as described above and in subclasses herein, wherein whenm is 1; when X₁, X₂, X₃ and X₄ are each ═O; when R₂ is —CN; when R₄ andR₆ are each —CH₃; when R₅ and R₇ are each —OCH₃; when R₈ is H; and R₁ is—NH(C═O)R_(C), then R_(C) is not —C(OH)(Me)CH₂(C═O)Me;

v) compounds as described above and in subclasses herein, wherein when mis 1 and when R₂ is H; and R₁ is —NH(C═O)R_(C), then R_(C) is not—CH(Me)NH(C═O)O(CH₂)Ph;

vi) compounds as described above and in subclasses herein, wherein whenm is 0; R₂ is H; X₃ is H; and R₁ is —C(═O)R_(A), then R_(A) is not—O(allcyl). Alternatively, in certain other embodiments, when R₂ is H; mis 1; and R₁ is —OR_(A), then R_(A) is not —C(═O)R_(C), or —S(O)₂R_(C),wherein R_(C) is an alkyl moiety.

vii) m is 0 or 1;

viii) R₂ is CN, —SCN, ═O, OH, protected hydroxyl, H, or alkoxy;

ix) R₂ is hydrogen, hydroxyl, —CN or —SCN;

x) R₃ is hydrogen, a nitrogen protecting group, —CN, —CH₂CN, aliphaticor aryl;

xi) R₄ and R₆ are each alkyl;

xii) R₅ and R₇ are each alkyloxy or thioalkyl;

xiii) R₈ is hydrogen, alkyl, —OH, protected hydroxyl, ═O, —CN, —SCN,halogen, —SH, protected thio, alkoxy, thioalkyl, amino, protected amino,or alkylamino;

xiv) R₈ is hydrogen;

xv) X₁, X₂, X₃, and X₄ are each independently alkoxy, —OH, protectedhydroxyl, or ═O;

xvi) R₂ is CN, —SCN, ═O, OH, protected hydroxyl, H, or alkoxy; R₃ ishydrogen, a nitrogen protecting group, —CN, —CH₂CN, aliphatic, or aryl;R₄ and R₆ are each alkyl; R₅ and R₇ are each alkyloxy or thioalkyl; R₈is hydrogen, alkyl, —OH, protected hydroxyl, ═O, CN, halogen, SH,alkoxy, thioalkyl, amino, or alkylamino; and X₁, X₂, X₃, and X₄ are eachindependently alkoxy, OH or ═O;

xvii) R₂ is —CN, —SCN, —OH, protected hydroxyl, H, or alkoxy; R₃ ishydrogen, a nitrogen protecting group, aliphatic, or aryl; R₄ and R₆ areeach alkyl; R₅ and R₇ are each alkyloxy or thioalkyl; X₁ and X₄ are each—OH; R₈ is hydrogen, alkyl, OH, protected hydroxyl, ═O, CN, halogen, SH,alkoxy, thioalkyl, amino, or alkylamino; and X₂ and X₃ are each alkyloxyor thioalkyl;

xviii) X₁ is OH, X₂ is OCH₃, X₃ is OCH₃, X₄ is OH, R₂ is CN, H or OH, R₃is Me, R₄ is CH₃, R₅ is OCH₃, R₆ is CH₃, R₇ is OCH₃, and R₈ is H;

xix) R₁ is OR_(A), SR_(A), or NR_(A)R_(B), wherein R_(A) and R_(B) areeach independently hydrogen, —(C═O)R_(C) or an aliphatic,heteroaliphatic, aryl, or heteroaryl moiety, wherein R_(C) is—(C═O)R_(D), or an aliphatic, heteroaliphatic, aryl or heteroarylmoiety, and wherein R_(D) is an aliphatic, heteroaliphatic, aryl, orheteroaryl moiety, or wherein R_(A) and R_(B), taken together, form aheterocyclic moiety,

whereby each of said aliphatic and heteroaliphatic moieties isindependently substituted or unsubstituted, branched or unbranched, orcyclic or acyclic, and each of said aryl, heteroaryl and heterocyclicmoieties is independently substituted or unsubstituted;

xx) R₁ is OR_(A), SR_(A), or NR_(A)R_(B), wherein R_(A) and R_(B) areeach independently hydrogen, —(C═O)R_(C), or an aryl, (aliphatic)aryl,(heteroaliphatic)aryl, heteroaryl, (aliphatic)heteroaryl, or(heteroaliphatic)heteroaryl moiety, wherein R_(C) is an aryl,(aliphatic)aryl, (heteroaliphatic)aryl, heteroaryl,(aliphatic)heteroaryl, or (heteroaliphatic)heteroaryl moiety, or whereinR_(A) and R_(B) taken together form a heterocyclic moiety,

whereby each of said aliphatic and heteroaliphatic moieties isindependently substituted or unsubstituted, branched or unbranched, orcyclic or acyclic, and each of said aryl, heteroaryl and heterocyclicmoieties is independently substituted or unsubstituted;

xxi) R₁ is —NR_(A)C(═O)R_(C), wherein R_(A) is hydrogen or lower alkyl,and R_(C) is a substituted or unsubstituted, branched or unbranched,cyclic or acyclic aliphatic or heteroaliphatic moiety, or a substitutedor unsubstituted aryl or heteroaryl moiety, or wherein R_(A) and R_(C)taken together form a heterocyclic or heteroaryl moiety;

xxii) R₁ is NR_(A)C(═O)R_(C), wherein R_(A) is hydrogen or lower alkyl,and R_(C) is an aryl, (aliphatic)aryl, (aliphatic)heteroaryl,heteroaryl, (heteroaliphatic)aryl, or (heteroaliphatic)heteroarylmoiety, or wherein R_(A) and R_(C) taken together form a heterocyclic orheteroaryl moiety;

whereby each of said aliphatic and heteroaliphatic moieties isindependently substituted or unsubstituted, branched or unbranched, orcyclic or acyclic, and each of said aryl, heteroaryl and heterocyclicmoieties is independently substituted or unsubstituted;

xxiii) R₁ is a substituted or unsubstituted, branched or unbranched,cyclic or acyclic aliphatic or heteroaliphatic moiety, or a substitutedor unsubstituted aryl or heteroaryl moiety;

xxiv) R₁ is an aryl, (aliphatic)aryl, (aliphatic)heteroaryl, heteroaryl,(heteroaliphatic)aryl, or (heteroaliphatic)heteroaryl moiety;

-   -   whereby each of said aliphatic and heteroaliphatic moieties is        independently substituted or unsubstituted, branched or        unbranched, or cyclic or acyclic, and each of said aryl,        heteroaryl and heterocyclic moieties is independently        substituted or unsubstituted;

xxv) any one or more of R₁, R_(A), R_(B), R_(C), or R_(D) isindependently any one of the following groups:

wherein each occurrence of R_(J) is independently hydrogen, a protectinggroup, —OR_(K), ═O, —C(═O)R_(K), —CO₂R_(K), —CN, —SCN, halogen, —SR_(K),—SOR_(K), —SO₂R_(K), —NO₂, —N(RK)₂, —NHC(O)R_(K), —B(ORK)₂, or analiphatic, heteroaliphatic, aryl, or heteroaryl moiety, wherein eachoccurrence of R_(K) is independently hydrogen, or an aliphatic,heteroaliphatic, aryl, or heteroaryl moiety, or wherein two occurrencesof R_(K), taken together form a cyclic aliphatic or heteroaliphaticmoiety; wherein each occurrence of Y is independently O, S or NH;wherein each occurrence of x is independently 0-5; and wherein eachoccurrence of n is independently 0-3, or wherein R_(J) is a labelingreagent,

whereby each of said aliphatic and heteroaliphatic moieties areindependently substituted or unsubstituted, branched or unbranched orcyclic or acyclic, and each of said aryl and heteroaryl moieties isindependently substituted or unsubstituted;

xxvi) R_(A) is hydrogen, R_(B) is —(C═O)R_(C), and R_(C) is iii, iv,vii, viii, ix, x, xv, or xvii, or R_(A) and R_(C) taken together formthe structure of i or ii;

xxvii) R_(A) is hydrogen, R_(B) is —(C═O)R_(C), and R_(C) is

xxviii) R_(J) is hydrogen, halogen, —OH, lower alkyl or lower alkoxy;

xxix) R_(J) is a linker-biotin or a linker-fluorescein moiety; and

xxx) x is 1 or 2.

As the reader will appreciate, compounds of particular interest include,among others, those which share the attributes of one or more of theforegoing subclasses. Some of those subclasses are illustrated by thefollowing sorts of compounds:I) Compounds of the Formula:

as described generally above and in classes and subclasses herein.

In certain embodiments, for compounds as described directly above, R_(A)is hydrogen, m is 1 and R_(C) is an aryl, (aliphatic)aryl,(aliphatic)heteroaryl, heteroaryl, (heteroaliphatic)aryl, or(heteroaliphatic)heteroaryl moiety, or wherein R_(A) and R_(C) takentogether form a heterocyclic or heteroaryl moiety, whereby each of saidaliphatic and heteroaliphatic moieties is independently substituted orunsubstituted, branched or unbranched, or cyclic or acyclic, and each ofsaid aryl, heteroaryl and heterocyclic moieties is independentlysubstituted or unsubstituted.

In certain other embodiments for compounds as described directly aboveR_(A) is hydrogen and R_(C) is any one of groups iii-xviii as describedherein, or R_(A) and R_(C) taken together are a structure of group i orii.

In still other embodiments, for compounds as described directly above,X₁ is OH, X₂ is OCH₃, X₃ is OCH₃, X₄ is OH, R₂ is CN, H or OH, R₃ is Me,R₄ is CH₃, R₅ is OCH₃, R₆ is CH₃, R₇ is OCH₃, R₈ is H, R_(A) ishydrogen, m is 1 and R_(C) is an aryl, (aliphatic)aryl,(aliphatic)heteroaryl, heteroaryl, (heteroaliphatic)aryl, or(heteroaliphatic)heteroaryl moiety, or wherein R_(A) and R_(C) takentogether form a heterocyclic or heteroaryl moiety, whereby each of saidaliphatic and heteroaliphatic moieties is independently substituted orunsubstituted, branched or unbranched, or cyclic or acyclic, and each ofsaid aryl, heteroaryl and heterocyclic moieties is independentlysubstituted or unsubstituted.

In yet other embodiments, for compounds as described directly above, X₁is OH, X₂ is OCH₃, X₃ is OCH₃, X₄ is OH, R₂ is CN, H or OH, R₃ is Me, R₄is CH₃, R₅ is OCH₃, R₆ is CH₃, R₇ is OCH₃, R₈ is H, R_(A) is hydrogenand R_(C) is any one of groups iii-xviii as described herein, or R_(A)and R_(C) taken together are a structure of group i or ii.II) Compounds of the Formula:

as described generally above and in classes and subclasses herein.

In certain embodiments, for compounds as described directly above, R_(A)is hydrogen, m is 1 and R_(C) is an aryl, (aliphatic)aryl,(aliphatic)heteroaryl, heteroaryl, (heteroaliphatic)aryl, or(heteroaliphatic)heteroaryl moiety, or wherein R_(A) and R_(C) takentogether form a heterocyclic or heteroaryl moiety, whereby each of saidaliphatic and heteroaliphatic moieties is independently substituted orunsubstituted, branched or unbranched, or cyclic or acyclic, and each ofsaid aryl, heteroaryl and heterocyclic moieties is independentlysubstituted or unsubstituted.

In certain other embodiments for compounds as described directly aboveR_(A) is hydrogen and R_(C) is any one of groups iii-xviii as describedherein, or R_(A) and R_(C) taken together are a structure of group i orii.

In still other embodiments, for compounds as described directly above,X₁ is OH, X₂ is OCH₃, X₃ is OCH₃, X is OH, R₂ is CN, H or OH, R₃ is Me,R₄ is CH₃, R₅ is OCH₃, R₆ is CH₃, R₇ is OCH₃, R₈ is H, R_(A) ishydrogen, m is 1 and R_(C) is an aryl, (aliphatic)aryl,(aliphatic)heteroaryl, heteroaryl, (heteroaliphatic)aryl, or(heteroaliphatic)heteroaryl moiety, or wherein R_(A) and R_(C) takentogether form a heterocyclic or heteroaryl moiety, whereby each of saidaliphatic and heteroaliphatic moieties is independently substituted orunsubstituted, branched or unbranched, or cyclic or acyclic, and each ofsaid aryl, heteroaryl and heterocyclic moieties is independentlysubstituted or unsubstituted.

In yet other embodiments, for compounds as described directly above, X₁is OH, X₂ is OCH₃, X₃ is OCH₃, X₄ is OH, R₂ is CN, H or OH, R₃ is Me, R₄is CH₃, R₅ is OCH₃, R₆ is CH₃, R₇ is OCH₃, R₈ is H, R_(A) is hydrogenand R_(C) is any one of groups iii-xviii as described herein, or R_(A)and R_(C) taken together are a structure of group i or ii.III) Compounds of the Formula:

as described generally above and in classes and subclasses herein.

In certain embodiments, for compounds as described directly above, m is1 and R_(A) is an aryl, (aliphatic)aryl, (aliphatic)heteroaryl,heteroaryl, (heteroaliphatic)aryl, or (heteroaliphatic)heteroarylmoiety, whereby each of said aliphatic and heteroaliphatic moieties isindependently substituted or unsubstituted, branched or unbranched, orcyclic or acyclic, and each of said aryl, heteroaryl and heterocyclicmoieties is independently substituted or unsubstituted.

In certain other embodiments for compounds as described directly aboveR_(A) is any one of groups iii-xviii as described herein.

In still other embodiments, for compounds as described directly above,X₁ is OH, X₂ is OCH₃, X₃ is OCH₃, X₄ is OH, R₂ is CN, H or OH, R₃ is Me,R₄ is CH₃, R₅ is OCH₃, R₆ is CH₃, R₇ is OCH₃, R₈ is H, m is 1 and R_(A)is an aryl, (aliphatic)aryl, (aliphatic)heteroaryl, heteroaryl,(heteroaliphatic)aryl, or (heteroaliphatic)heteroaryl moiety, wherebyeach of said aliphatic and heteroaliphatic moieties is independentlysubstituted or unsubstituted, branched or unbranched, or cyclic oracyclic, and each of said aryl, heteroaryl and heterocyclic moieties isindependently substituted or unsubstituted.

In yet other embodiments, for compounds as described directly above, X₁is OH, X₂ is OCH₃, X₃ is OCH₃, X₄ is OH, R₂ is CN, H or OH, R₃ is Me, R₄is CH₃, R₅ is OCH₃, R₆ is CH₃, R₇ is OCH₃, R₈ is H, R_(A) is any one ofgroups iii-xviii as described herein.IV) Compounds of the Formula:

as described generally above and in classes and subclasses herein.

In certain embodiments, for compounds as described directly above, m is1 and R_(A) is an aryl, (aliphatic)aryl, (aliphatic)heteroaryl,heteroaryl, (heteroaliphatic)aryl, or (heteroaliphatic)heteroarylmoiety, whereby each of said aliphatic and heteroaliphatic moieties isindependently substituted or unsubstituted, branched or unbranched, orcyclic or acyclic, and each of said aryl, heteroaryl and heterocyclicmoieties is independently substituted or unsubstituted.

In certain other embodiments for compounds as described directly aboveR_(A) is any one of groups iii-xviii as described herein.

In still other embodiments, for compounds as described directly above,X₁ is OH, X₂ is OCH₃, X₃ is OCH₃, X₄ is OH, R₂ is CN, H or OH, R₃ is Me,R₄ is CH₃, R₅ is OCH₃, R₆ is CH₃, R₇ is OCH₃, R₈ is H, m is 1 and R_(A)is an aryl, (aliphatic)aryl, (aliphatic)heteroaryl, heteroaryl,(heteroaliphatic)aryl, or (heteroaliphatic)heteroaryl moiety, wherebyeach of said aliphatic and heteroaliphatic moieties is independentlysubstituted or unsubstituted, branched or unbranched, or cyclic oracyclic, and each of said aryl, heteroaryl and heterocyclic moieties isindependently substituted or unsubstituted.

In yet other embodiments, for compounds as described directly above, X₁is OH, X₂ is OCH₃, X₃ is OCH₃, X₄ is OH, R₂ is CN, H or OH, R₃ is Me, R₄is CH₃, R₅ is OCH₃, R₆ is CH₃, R₇ is OCH₃, R₈ is H, R_(A) is any one ofgroups iii-xviii as described herein.V. Compounds of the Formula:

as described generally above and in classes and subclasses herein.

In certain embodiments, for compounds as described directly above, m is1 and R_(A) is an aryl, (aliphatic)aryl, (aliphatic)heteroaryl,heteroaryl, (heteroaliphatic)aryl, or (heteroaliphatic)heteroarylmoiety, whereby each of said aliphatic and heteroaliphatic moieties isindependently substituted or unsubstituted, branched or unbranched, orcyclic or acyclic, and each of said aryl, heteroaryl and heterocyclicmoieties is independently substituted or unsubstituted.

In certain other embodiments for compounds as described directly aboveR_(A) is any one of groups iii-xviii as described herein.

In still other embodiments, for compounds as described directly above,X₁ is OH, X₂ is OCH₃, X₃ is OCH₃, X₄ is OH, R₂ is CN, H or OH, R₃ is Me,R₄ is CH₃, R₅ is OCH₃, R₆ is CH₃, R₇ is OCH₃, R₈ is H, m is 1 and R_(A)is an aryl, (aliphatic)aryl, (aliphatic)heteroaryl, heteroaryl,(heteroaliphatic)aryl, or (heteroaliphatic)heteroaryl moiety, wherebyeach of said aliphatic and heteroaliphatic moieties is independentlysubstituted or unsubstituted, branched or unbranched, or cyclic oracyclic, and each of said aryl, heteroaryl and heterocyclic moieties isindependently substituted or unsubstituted.

In yet other embodiments, for compounds as described directly above, X₁is OH, X₂ is OCH₃, X₃ is OCH₃, X₄ is OH, R₂ is CN, H or OH, R₃ is Me, R₄is CH₃, R₅ is OCH₃, R₆ is CH₃, R₇ is OCH₃, R₈ is H, R_(A) is any one ofgroups iii-xviii as described herein.VI. Compounds of the Formula:

as described generally above and in classes and subclasses herein.

In certain embodiments, for compounds as described directly above, m is1 and R_(A) is an aryl, (aliphatic)aryl, (aliphatic)heteroaryl,heteroaryl, (heteroaliphatic)aryl, or (heteroaliphatic)heteroarylmoiety, whereby each of said aliphatic and heteroaliphatic moieties isindependently substituted or unsubstituted, branched or unbranched, orcyclic or acyclic, and each of said aryl, heteroaryl and heterocyclicmoieties is independently substituted or unsubstituted.

In certain other embodiments for compounds as described directly aboveR_(A) is any one of groups iii-xviii as described herein.

In still other embodiments, for compounds as described directly above,X₁ is OH, X₂ is OCH₃, X₃ is OCH₃, X₄ is OH, R₂ is CN, H or OH, R₃ is Me,R₄ is CH₃, R₅ is OCH₃, R₆ is CH₃, R₇ is OCH₃, R₈ is H, m is 1 and R_(A)is an aryl, (aliphatic)aryl, (aliphatic)heteroaryl, heteroaryl,heteroaliphatic)aryl, or (heteroaliphatic)heteroaryl moiety, wherebyeach of said aliphatic and heteroaliphatic moieties is independentlysubstituted or unsubstituted, branched or unbranched, or cyclic oracyclic, and each of said aryl, heteroaryl and heterocyclic moieties isindependently substituted or unsubstituted.

In yet other embodiments, for compounds as described directly above, X₁is OH, X₂ is OCH₃, X₃ is OCH₃, X₄ is OH, R₂ is CN, H or OH, R₃ is Me, R₄is CH₃, R₅ is OCH₃, R₆ is CH₃, R₇ is OCH₃, R₈ is H, R_(A) is any one ofgroups iii-xviii as described herein.VII. Compounds of the Formula:

wherein R₁ is an aliphatic, heteroaliphatic, aryl, (aliphatic)aryl,(aliphatic)heteroaryl, heteroaryl, (heteroaliphatic)aryl, or(heteroaliphatic)heteroaryl moiety, whereby each of said aliphatic andheteroaliphatic moieties is independently substituted or unsubstituted,branched or unbranched, or cyclic or acyclic, and each of said aryl,heteroaryl and heterocyclic moieties is independently substituted orunsubstituted.

In certain other embodiments for compounds as described directly aboveR₁ is any one of groups iii-xviii as described herein.

In still other embodiments, for compounds as described directly above,X₁ is OH, X₂ is OCH₃, X₃ is OCH₃, X₄ is OH, R₂ is CN, H or OH, R₃ is Me,R₄ is CH₃, R₅ is OCH₃, R₆ is CH₃, R₇ is OCH₃, R₈ is H, m is 0 or 1 andR₁ is an aliphatic, heteroaliphatic, aryl, (aliphatic)aryl,(aliphatic)heteroaryl, heteroaryl, (heteroaliphatic)aryl, or(heteroaliphatic)heteroaryl moiety, whereby each of said aliphatic andheteroaliphatic moieties is independently substituted or unsubstituted,branched or unbranched, or cyclic or acyclic, and each of said aryl,heteroaryl and heterocyclic moieties is independently substituted orunsubstituted.

In yet other embodiments, for compounds as described directly above, X₁is OH, X₂ is OCH₃, X₃ is OCH₃, X₄ is OH, R₂ is CN, H or OH, R₃ is Me, R₄is CH₃, R₅ is OCH₃, R₆ is CH₃, R₇ is OCH₃, R₈ is H, R₁ is any one ofgroups iii-xviii as described herein.VIII. Compounds of the Formula:

wherein R₁ is an aliphatic, heteroaliphatic, aryl, (aliphatic)aryl,(aliphatic)heteroaryl, heteroaryl, (heteroaliphatic)aryl, orheteroaliphatic)heteroaryl moiety, whereby each of said aliphatic andheteroaliphatic moieties is independently substituted or unsubstituted,branched or unbranched, or cyclic or acyclic, and each of said aryl,heteroaryl and heterocyclic moieties is independently substituted orunsubstituted.

In certain other embodiments for compounds as described directly aboveR₁ is any one of groups iii-xviii as described herein.

In still other embodiments, for compounds as described directly above,X₁ is OH, X₂ is OCH₃, X₃ is OCH₃, X₄ is OH, R₂ is CN, H or OH, R₃ is Me,R₄ is CH₃, R₅ is OCH₃, R₆ is CH₃, R₇ is OCH₃, R₈ is H, m is 0 or 1 andR₁ is an aliphatic, heteroaliphatic, aryl, (aliphatic)aryl,(aliphatic)heteroaryl, heteroaryl, (heteroaliphatic)aryl, or(heteroaliphatic)heteroaryl moiety, whereby each of said aliphatic andheteroaliphatic moieties is independently substituted or unsubstituted,branched or unbranched, or cyclic or acyclic, and each of said aryl,heteroaryl and heterocyclic moieties is independently substituted orunsubstituted.

In yet other embodiments, for compounds as described directly above, X₁is OH, X₂ is OCH₃, X₃ is OCH₃, X₄ is OH, R₂ is CN, H or OH, R₃ is Me, R₄is CH₃, R₅ is OCH₃, R₆ is CH₃, R₇ is OCH₃, R₈ is H, R₁ is any one ofgroups iii-xviii as described herein.

It will be appreciated that some of the foregoing classes and subclassesof compounds can exist in various isomeric forms. The inventionencompasses the compounds as individual isomers substantially free ofother isomers and alternatively, as mixtures of various isomers, e.g.,racemic mixtures of stereoisomers. Additionally, the inventionencompasses both (Z) and (E) double bond isomers unless otherwisespecifically designated. The invention also encompasses tautomers ofspecific compounds as described above. In addition to theabove-mentioned compounds per se, this invention also encompassespharmaceutically acceptable derivatives of these compounds andcompositions comprising one or more compounds of the invention and oneor more pharmaceutically acceptable excipients or additives.

Compounds of this invention which are of particular interest includethose which:

-   -   exhibit cytotoxic or growth inhibitory effect on cancer cell        lines maintained in vitro or in animal studies using a        scientifically acceptable cancer cell xenograft model;    -   exhibit enhanced water solubility over existing chemotherapetuic        agents, or additionally or alternatively exhibit sufficient        solubility to be formulated in an aqueous medium; and    -   exhibit a therapeutic profile (e.g., optimum safety and curative        effect) that is superior to existing chemotherapeutic agents.

This invention also provides a pharmaceutical preparation comprising atleast one of the compounds as described above and herein, or apharmaceutically acceptable derivative thereof, which compounds arecapable of inhibiting the growth of or killing cancer cells, and, incertain embodiments of special interest are capable of inhibiting thegrowth of or killing multidrug resistant cancer cells.

The invention further provides a method for inhibiting tumor growthand/or tumor metastasis. In certain embodiments of special interest, theinvention provides a method of treating cancers by inhibiting tumorgrowth and/or tumor metastasis for tumors multidrug resistant cancercells. The method involves the administration of a therapeuticallyeffective amount of the compound or a pharmaceutically acceptablederivative thereof to a subject (including, but not limited to a humanor animal) in need of it. In certain embodiments, specifically fortreating cancers comprising multidrug resistant cancer cells, thetherapeutically effective amount is an amount sufficient to kill orinhibit the growth of multidrug resistant cancer cell lines. In certainembodiments, the inventive compounds are useful for the treatment ofsolid tumors.

3) Compounds and Definitions

As discussed above, the present invention provides a novel class ofcompounds useful for the treatment of cancer and other proliferativeconditions related thereto. Compounds of this invention comprise those,as set forth above and described herein, and are illustrated in part bythe various classes, subgenera and species disclosed elsewhere herein.

It will be appreciated by one of ordinary skill in the art that numerousasymmetric centers exist in the compounds of the present invention.Thus, inventive compounds and pharmaceutical compositions thereof may bein the form of an individual enantiomer, diastereomer or geometricisomer, or may be in the form of a mixture of stereoisomers.Additionally, in certain embodiments, as detailed herein, the method ofthe present invention provides for the stereoselective synthesis ofalkaloids and analogues thereof. Thus, in certain embodiments, thecompounds of the invention are enantiopure.

Additionally, the present invention provides pharmaceutically acceptablederivatives of the inventive compounds, and methods of treating asubject using these compounds, pharmaceutical compositions thereof, oreither of these in combination with one or more additional therapeuticagents. The phrase, “pharmaceutically acceptable derivative”, as usedherein, denotes any pharmaceutically acceptable salt, ester, or salt ofsuch ester, of such compound, or any other adduct or derivative which,upon administration to a patient, is capable of providing (directly orindirectly) a compound as otherwise described herein, or a metabolite orresidue thereof. Pharmaceutically acceptable derivatives thus includeamong others pro-drugs. A pro-drug is a derivative of a compound,usually with significantly reduced pharmacological activity, whichcontains an additional moiety that is susceptible to removal in vivoyielding the parent molecule as the pharmacologically active species. Anexample of a pro-drug is an ester that is cleaved in vivo to yield acompound of interest. Pro-drugs of a variety of compounds, and materialsand methods for derivatizing the parent compounds to create thepro-drugs, are known and may be adapted to the present invention.Certain exemplary pharmaceutical compositions and pharmaceuticallyacceptable derivatives will be discussed in more detail herein below.

Certain compounds of the present invention, and definitions of specificfunctional groups are also described in more detail below. For purposesof this invention, the chemical elements are identified in accordancewith the Periodic Table of the Elements, CAS version, Handbook ofChemistry and Physics, 75^(th) Ed., inside cover, and specificfunctional groups are generally defined as described therein.Additionally, general principles of organic chemistry, as well asspecific functional moieties and reactivity, are described in “OrganicChemistry”, Thomas Sorrell, University Science Books, Sausalito: 1999,the entire contents of which are incorporated herein by reference.Furthermore, it will be appreciated by one of ordinary skill in the artthat the synthetic methods, as described herein, utilize a variety ofprotecting groups. By the term “protecting group”, has used herein, itis meant that a particular functional moiety, e.g., C, O, S, or N, istemporarily blocked so that a reaction can be carried out selectively atanother reactive site in a multifunctional compound. In preferredembodiments, a protecting group reacts selectively in good yield to givea protected substrate that is stable to the projected reactions; theprotecting group must be selectively removed in good yield by readilyavailable, preferably nontoxic reagents that do not attack the otherfunctional groups; the protecting group forms an easily separablederivative (more preferably without the generation of new stereogeniccenters); and the protecting group has a minimum of additionalfunctionality to avoid further sites of reaction. As detailed herein,oxygen, sulfur, nitrogen and carbon protecting groups may be utilized.Exemplary protecting groups are detailed herein, however, it will beappreciated that the present invention is not intended to be limited tothese protecting groups; rather, a variety of additional equivalentprotecting groups can be readily identified using the above criteria andutilized in the method of the present invention. Additionally, a varietyof protecting groups are described in “Protective Groups in OrganicSynthesis” Third Ed. Greene, T. W. and Wuts, P. G., Eds., John Wiley &Sons, New York: 1999, the entire contents of which are herebyincorporated by reference. Furthermore, a variety of carbon protectinggroups are described in Myers, A.; Kung, D. W.; Zhong, B.; Movassaghi,M.; Kwon, S. J. Am. Chem. Soc. 1999, 121, 8401-8402, the entire contentsof which are hereby incorporated by reference.

It will be appreciated that the compounds, as described herein, may besubstituted with any number of substituents or functional moieties. Ingeneral, the term “substituted” whether preceded by the term“optionally” or not, and substituents contained in formulas of thisinvention, refer to the replacement of hydrogen radicals in a givenstructure with the radical of a specified substituent. When more thanone position in any given structure may be substituted with more thanone substituent selected from a specified group, the substituent may beeither the same or different at every position. As used herein, the term“substituted” is contemplated to include all permissible substituents oforganic compounds. In a broad aspect, the permissible substituentsinclude acyclic and cyclic, branched and unbranched, carbocyclic andheterocyclic, aromatic and nonaromatic substituents of organiccompounds. For purposes of this invention, heteroatoms such as nitrogenmay have hydrogen substituents and/or any permissible substituents oforganic compounds described herein which satisfy the valencies of theheteroatoms. Furthermore, this invention is not intended to be limitedin any manner by the permissible substituents of organic compounds.Combinations of substituents and variables envisioned by this inventionare preferably those that result in the formation of stable compoundsuseful in the treatment, for example of proliferative disorders,including, but not limited to cancer. The term “stable”, as used herein,preferably refers to compounds which possess stability sufficient toallow manufacture and which maintain the integrity of the compound for asufficient period of time to be detected and preferably for a sufficientperiod of time to be useful for the purposes detailed herein.

The term “acyl”, as used herein, refers to a carbonyl-containingfunctionality, e.g., —C(═O)R_(x), wherein R_(x) is an aliphatic,heteroaliphatic, aryl, heteroaryl, (aliphatic)aryl,(heteroaliphatic)aryl, heteroaliphatic(aryl) orheteroaliphatic(heteroaryl) moiety, whereby each of the aliphatic,heteroaliphatic, aryl, or heteroaryl moieties is substituted orunsubstituted, or is a substituted (e.g., hydrogen or aliphatic,heteroaliphatic, aryl, or heteroaryl moieties) oxygen or nitrogencontaining functionality (e.g., forming a carboxylic acid, ester, oramide functionality).

The term “aliphatic”, as used herein, includes both saturated andunsaturated, straight chain (i.e., unbranched), branched, cyclic, orpolycyclic aliphatic hydrocarbons, which are optionally substituted withone or more functional groups. As will be appreciated by one of ordinaryskill in the art, “aliphatic” is intended herein to include, but is notlimited to, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, andcycloalkynyl moieties. Thus, as used herein, the term “alkyl” includesstraight, branched and cyclic alkyl groups. An analogous conventionapplies to other generic terms such as “alkenyl”, “alkynyl” and thelike. Furthermore, as used herein, the terms “alkyl”, “alkenyl”,“alkynyl” and the like encompass both substituted and unsubstitutedgroups. In certain embodiments, as used herein, “lower alkyl” is used toindicate those alkyl groups (cyclic, acyclic, substituted,unsubstituted, branched or unbranched) having 1-6 carbon atoms.

In certain embodiments, the alkyl, alkenyl and alkynyl groups employedin the invention contain 1-20 aliphatic carbon atoms. In certain otherembodiments, the alky, alkenyl, and alkynyl groups employed in theinvention contain 1-10 aliphatic carbon atoms. In yet other embodiments,the alkyl, alkenyl, and alkynyl groups employed in the invention contain1-8 aliphatic carbon atoms. In still other embodiments, the alkyl,alkenyl, and alkynyl groups employed in the invention contain 1-6aliphatic carbon atoms. In yet other embodiments, the alkyl, alkenyl,and alkynyl groups employed in the invention contain 1-4 carbon atoms.Illustrative aliphatic groups thus include, but are not limited to, forexample, methyl, ethyl, n-propyl, isopropyl, cyclopropyl,—CH₂-cyclopropyl, allyl, n-butyl, sec-butyl, isobutyl, tert-butyl,cyclobutyl, —CH₂-cyclobutyl, n-pentyl, sec-pentyl, isopentyl,tert-pentyl, cyclopentyl, —CH₂-cyclopentyl, n-hexyl, sec-hexyl,cyclohexyl, —CH₂-cyclohexyl moieties and the like, which again, may bearone or more substituents. Alkenyl groups include, but are not limitedto, for example, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl, andthe like. Representative alkynyl groups include, but are not limited to,ethynyl, 2-propynyl (propargyl), 1-propynyl and the like.

The term “alkoxy”, or “thioalkyl” as used herein refers to an alkylgroup, as previously defined, attached to the parent molecular moietythrough an oxygen atom or through a sulfur atom. In certain embodiments,the alkyl group contains 1-20 aliphatic carbon atoms. In certain otherembodiments, the alkyl group contains 1-10 aliphatic carbon atoms. Inyet other embodiments, the alkyl, alkenyl, and alkynyl groups employedin the invention contain 1-8 aliphatic carbon atoms. In still otherembodiments, the alkyl group contains 1-6 aliphatic carbon atoms. In yetother embodiments, the alkyl group contains 14 aliphatic carbon atoms.Examples of alkoxy, include but are not limited to, methoxy, ethoxy,propoxy, isopropoxy, n-butoxy, tert-butoxy, neopentoxy and n-hexoxy.Examples of thioalkyl include, but are not limited to, methylthio,ethylthio, propylthio, isopropylthio, n-butylthio, and the like.

The term “alkylamino” refers to a group having the structure —NHR′wherein R′ is alkyl, as defined herein. In certain embodiments, thealkyl group contains 1-20 aliphatic carbon atoms. In certain otherembodiments, the alkyl group contains 1-10 aliphatic carbon atoms. Inyet other embodiments, the alkyl, alkenyl, and alkynyl groups employedin the invention contain 1-8 aliphatic carbon atoms. In still otherembodiments, the alkyl group contains 1-6 aliphatic carbon atoms. In yetother embodiments, the alkyl group contains 14 aliphatic carbon atoms.Examples of alkylamino include, but are not limited to, methylamino,ethylamino, iso-propylamino and the like.

Some examples of substituents of the above-described aliphatic (andother) moieties of compounds of the invention include, but are notlimited to aliphatic; heteroaliphatic; aryl; heteroaryl; alkylaryl;alkylheteroaryl; alkoxy; aryloxy; heteroalkoxy; heteroaryloxy;alkylthio; arylthio; heteroalkylthio; heteroarylthio; F; Cl; Br; I; —OH;—NO₂; —CN; —SCN; —CF₃; —CH₂CF₃; —CHCl₂; —CH₂OH; CH₂CH₂OH; —CH₂NH₂;—CH₂SO₂CH₃; —C(O)R_(x); —CO₂(R_(x)); —CON(R_(x))₂; —OC(O)R_(x);—OCO₂R_(x); —OCON(R_(x))₂; —N(R_(x)); —S(O)₂R_(x); —NR_(x)(CO)R_(x) or—B(OR_(x))₂, wherein each occurrence of R_(x) independently includes,but is not limited to, aliphatic, heteroaliphatic, aryl, heteroaryl,alkylaryl, or alkylheteroaryl, or wherein any two of R_(x), takentogether is a cyclic aliphatic, heteroaliphatic, aryl or heteroarylmoiety, wherein any of the aliphatic, heteroaliphatic, alkylaryl, oralkylheteroaryl substituents described above and herein may besubstituted or unsubstituted, branched or unbranched, cyclic or acyclic,and wherein any of the aryl or heteroaryl substituents described aboveand herein may be substituted or unsubstituted. Additional examples ofgenerally applicable substituents are illustrated by the specificembodiments shown in the Examples that are described herein.

In general, the terms “aryl” and “heteroaryl”, as used herein, refer tostable mono- or polycyclic, heterocyclic, polycyclic, andpolyheterocyclic unsaturated moieties having preferably 3-14 carbonatoms, each of which may be substituted or unsubstituted. Substituentsinclude, but are not limited to, any of the previously mentionedsubstitutents, i.e., the substituents recited for aliphatic moieties, orfor other moieties as disclosed herein, resulting in the formation of astable compound. In certain embodiments of the present invention, “aryl”refers to a mono- or bicyclic carbocyclic ring system having one or twoaromatic rings including, but not limited to, phenyl, naphthyl,tetrahydronaphthyl, indanyl, indenyl and the like. In certainembodiments of the present invention, the term “heteroaryl”, as usedherein, refers to a cyclic aromatic radical having from five to ten ringatoms of which one ring atom is selected from S, O and N; zero, one ortwo ring atoms are additional heteroatoms independently selected from S,O and N; and the remaining ring atoms are carbon, the radical beingjoined to the rest of the molecule via any of the ring atoms, such as,for example, pyridyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl,imidazolyl, thiazolyl, oxazolyl, isooxazolyl, thiadiazolyl,oxadiazolyl,thiophenyl, furanyl, quinolinyl, isoquinolinyl, and the like.

It will be appreciated that aryl and heteroaryl groups (includingbicyclic aryl groups) can be unsubstituted or substituted, whereinsubstitution includes replacement of one, two or three of the hydrogenatoms thereon independently with any one or more of the followingmoieties including, but not limited to: aliphatic; heteroaliphatic;aryl; heteroaryl; alkylaryl; alkylheteroaryl; alkoxy; aryloxy;heteroalkoxy; heteroaryloxy; alkylthio; arylthio; heteroalkylthio;heteroarylthio; F; Cl; Br; I; —OH; —NO₂; —CN; —SCN; —CF₃; —CH₂CF₃;—CHCl₂; —CH₂OH; —CH₂CH₂OH; —CH₂NH₂; —CH₂SO₂CH₃; —C(O)R_(x); —CO₂(R_(x));—CON(R_(x))₂; —OC(O)R_(x); —OCO₂R_(x); —OCON(R_(x))₂; —N(R_(x))₂;—S(O)₂R_(x); —NR_(x)(CO)R_(x) or —B(OR_(x))₂, wherein each occurrence ofR_(x) independently includes, but is not limited to, aliphatic,heteroaliphatic, aryl, heteroaryl, alkylaryl, or alkylheteroaryl, orwherein any two of R_(x), taken together is a cyclic aliphatic,heteroaliphatic, aryl or heteroaryl moiety, wherein any of thealiphatic, heteroaliphatic, alkylaryl, or alkylheteroaryl substituentsdescribed above and herein may be substituted or unsubstituted, branchedor unbranched, cyclic or acyclic, and wherein any of the aryl orheteroaryl substituents described above and herein may be substituted orunsubstituted. Additional examples of generally applicable substituentsare illustrated by the specific embodiments shown in the Examples thatare described herein.

The term “cycloalkyl”, as used herein, refers specifically to groupshaving three to seven, preferably three to ten carbon atoms. Suitablecycloalkyls include, but are not limited to cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl and the like, which, as in the caseof other aliphatic, heteroaliphatic or hetercyclic moieties, mayoptionally be substituted with substituents including, but not limitedto aliphatic; heteroaliphatic; aryl; heteroaryl; alkylaryl;alkylheteroaryl; alkoxy; aryloxy; heteroalkoxy; heteroaryloxy;alkylthio; arylthio; heteroalkylthio; heteroarylthio; F; Cl; Br; I; —OH;—NO₂; —CN; —SCN; —CF₃; —CH₂CF₃; —CHCl₂; —CH₂OH; —CH₂CH₂OH; —CH₂NH₂;—CH₂SO₂CH₃; —C(O)R_(x); —CO₂(R_(x)); —CON(R_(x))₂; —OC(O)R_(x);—OCO₂R_(x); —OCON(R_(x))₂; —N(R_(x))₂; —S(O)₂R_(x); —NR_(x)(CO)R_(x) or—B(OR_(x))₂, wherein each occurrence of R_(x) independently includes,but is not limited to, aliphatic, heteroaliphatic, aryl, heteroaryl,alkylaryl, or alkylheteroaryl, or wherein any two of R_(x), takentogether is a cyclic aliphatic, heteroaliphatic, aryl or heteroarylmoiety, wherein any of the aliphatic, heteroaliphatic, alkylaryl, oralkylheteroaryl substituents described above and herein may besubstituted or unsubstituted, branched or unbranched, cyclic or acyclic,and wherein any of the aryl or heteroaryl substituents described aboveand herein may be substituted or unsubstituted. Additional examples ofgenerally applicable substituents are illustrated by the specificembodiments shown in the Examples that are described herein.

The term “heteroaliphatic”, as used herein, refers to aliphatic moietiesthat contain one or more oxygen, sulfur, nitrogen, phosphorus or siliconatoms, e.g., in place of carbon atoms. Heteroaliphatic moieties may bebranched, unbranched, cyclic or acyclic and include saturated andunsaturated heterocycles such as morpholino, pyrrolidinyl, etc. Incertain embodiments, heteroaliphatic moieties are substituted byindependent replacement of one or more of the hydrogen atoms thereonwith one or more moieties including, but not limited to aliphatic;heteroaliphatic; aryl; heteroaryl; alkylaryl; alkylheteroaryl; alkoxy;aryloxy; heteroalkoxy; heteroaryloxy; alkylthio; arylthio;heteroalkylthio; heteroarylthio; F; Cl; Br; I; —OH; —NO₂; —CN; —SCN;—CF₃; —CH₂CF₃; —CHCl₂; —CH₂OH; —CH₂CH₂OH; —CH₂NH₂; —CH₂SO₂CH₃;—C(O)R_(x); —CO₂(R_(x)); —CON(R_(x))₂; —OC(O)R_(x); —OCO₂R_(x);—OCON(R_(x))₂; —N(R_(x))₂; —S(O)₂R_(x); —NR_(x)(CO)R_(x) or —B(OR_(x))₂,wherein each occurrence of R_(x) independently includes, but is notlimited to, aliphatic, heteroaliphatic, aryl, heteroaryl, alkylaryl, oralkylheteroaryl, or wherein any two of R_(x), taken together is a cyclicaliphatic, heteroaliphatic, aryl or heteroaryl moiety, wherein any ofthe aliphatic, heteroaliphatic, alkylaryl, or alkylheteroarylsubstituents described above and herein may be substituted orunsubstituted, branched or unbranched, cyclic or acyclic, and whereinany of the aryl or heteroaryl substituents described above and hereinmay be substituted or unsubstituted. Additional examples of generallyapplicable substituents are illustrated by the specific embodimentsshown in the Examples that are described herein.

The terms “halo” and “halogen” as used herein refer to an atom selectedfrom fluorine, chlorine, bromine and iodine.

The term “haloalkyl” denotes an alkyl group, as defined above, havingone, two, or three halogen atoms attached thereto and is exemplified bysuch groups as chloromethyl, bromoethyl, trifluoromethyl, and the like.

The term “heterocycloalkyl” or “heterocycle”, as used herein, refers toa non-aromatic 5-, 6- or 7-membered ring or a polycyclic group,including, but not limited to a bi- or tri-cyclic group comprising fusedsix-membered rings having between one and three heteroatomsindependently selected from oxygen, sulfur and nitrogen, wherein (i)each 5-membered ring has 0 to 1 double bonds and each 6-membered ringhas 0 to 2 double bonds, (ii) the nitrogen and sulfur heteroatoms may beoptionally be oxidized, (iii) the nitrogen heteroatom may optionally bequaternized, and (iv) any of the above heterocyclic rings may be fusedto a benzene ring. Representative heterocycles include, but are notlimited to, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl,imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl,morpholinyl, thiazolidinyl, isothiazolidinyl, and tetrahydrofuryl. Incertain embodiments, a “substituted heterocycloalkyl or heterocycle”group is utilized and as used herein, refers to a heterocycloalkyl orheterocycle group, as defined above, substituted by the independentreplacement of one, two or three of the hydrogen atoms thereon with butare not limited to aliphatic; heteroaliphatic; aryl; heteroaryl;alkylaryl; alkylheteroaryl; alkoxy; aryloxy; heteroalkoxy;heteroaryloxy; alkylthio; arylthio; heteroalkylthio; heteroarylthio; F;Cl; Br; I; —OH; —NO₂; —CN; —SCN; —CF₃; —CH₂CF₃; —CHCl₂; —CH₂OH;—CH₂CH₂OH; —CH₂NH₂; —CH₂SO₂CH₃; —C(O)R_(x); —CO₂(R_(x)); —CON(R_(x))₂;—OC(O)R_(x); —OCO₂R_(x); —OCON(R_(x))₂; —N(R_(x))₂; —S(O)₂R_(x);—NR_(x)(CO)R_(x) or —B(OR_(x))₂, wherein each occurrence of R,independently includes, but is not limited to, aliphatic,heteroaliphatic, aryl, heteroaryl, alkylaryl, or alkylheteroaryl, orwherein any two of R_(x), taken together is a cyclic aliphatic,heteroaliphatic, aryl or heteroaryl moiety, wherein any of thealiphatic, heteroaliphatic, alkylaryl, or alkylheteroaryl substituentsdescribed above and herein may be substituted or unsubstituted, branchedor unbranched, cyclic or acyclic, and wherein any of the aryl orheteroaryl substituents described above and herein may be substituted orunsubstituted. Additional examples of generally applicable substituentsare illustrated by the specific embodiments shown in the Examples thatare described herein. “Labeled”: As used herein, the term “labeled” isintended to mean that a compound has at least one element, isotope orchemical compound attached to enable the detection of the compound. Itwill be appreciated that certain of the inventive compounds aresubstituted with a labeling reagent, that, as used herein, is intendedto mean an element, isotope or chemical compound attached to theinventive compound directly or through a suitable linker (e.g.,substituted or unsubstituted, branched or unbranched, cyclic or acyclic,aliphatic, heteroaliphatic, aryl, heteroaryl moiety). Certain exemplarylabeling reagents are described in the Exemplification herein; however,it will be appreciated that the present invention is not intended to belimited to these examples. Rather, a variety of labeling reagents can beemployed as substituents for the inventive compounds.

4. Synthetic Methodology:

It will be appreciated that each of the compounds as described above andherein can be synthesized according to the pioneering methodologydescribed in more detail herein; however, the synthesis of thesecompounds is not limited to the methodology described herein. Rather,any methods available in the art of synthetic organic chemistry can beutilized to provide the inventive compounds, including combinatorialtechniques. Significantly, the methodology as described herein (and asalso described in Myers, A. G., Plowright, A. T. J. Am Chem. Soc. 2001,123, 5114-5115; and Myers, A. G., Kung, D. W. J. Am. Chem. Soc. 1999,121, 10828-10829, the entire contents of which are hereby incorporatedby reference (including Supplemental Materials available via theinternet at http://publ.acs.org)) enables the efficient production ofsignificant quantities of alkaloid structures that can be utilizeddirectly or modified to generate other modified alkaloid structures.

In general, the pioneering method of the present invention provides ageneral method for the rapid synthesis of alkaloids comprising (1)providing a desired number of substituted aldehyde precursors; and (2)reacting said desired number of substituted aldehyde precursors undersuitable conditions to effect the directed condensation of said desiredsubstituted aldehyde precursors, whereby an alkaloid is generated. Itwill be appreciated that the alkaloid structures generated may, incertain embodiments, be natural product precursors, and thus thesubsequent reaction of said precursors with suitable reagents yields anatural product or derivatives thereof (e.g., saframycins or derivativesthereof). In certain other embodiments, the alkaloid structuresgenerated represent core structures that can subsequently befunctionalized to generate a variety of structures of interest forbiological testing and therapeutic use. In certain embodiments ofspecial interest, inventive compounds are prepared using solid phasemethodologies as described herein, for the efficient synthesis of largenumbers of exemplary compounds. In general, in certain exemplaryembodiments, the step of reacting said desired number of substitutedaldehyde precursors comprises reacting said precursors under conditionsto effect the directed condensation of said desired substituted aldehydeprecursors, whereby an alkaloid is generated. In certain embodiments,the substituted aldehyde precursors are α-amino aldehyde precursors. Incertain other embodiments, any combination of α-amino aldehyde and othersubstituted aldehyde precursors may be utilized (see, e.g.,Exemplification, Section IV) to diversify the core pentacyclicstructures.

In certain other embodiments of special interest, particularly thoserelated to the synthesis of saframycins and derivatives thereof, thestep of providing said desired number of precursors comprises providinga first N-protected precursor, a second C-protected precursor and athird substituted aldehyde precursor, and the step of reacting saidprecursors under suitable conditions further comprises: (1) reactingsaid first N-protected and said second C-protected aldehyde precursorunder suitable conditions to generate a tetrahydroisoquinoline corestructure; (2) reacting said third substituted aldehyde precursor withsaid tetrahydroisoquinoline core structure under suitable conditions togenerate a trimer of aldehydes; (3) reacting said trimer of aldehydesunder suitable conditions to generate an alkaloid; and (4) optionallyfurther reacting the alkaloid generated in step (3) to generate adiversified alkaloid. Additionally, in certain embodiments, thetetrahydroisoquinoline core structure generated in step (1) can bediversified prior to reaction in step (2) with the third aldehydeprecursor to generate additional diversity. In certain embodiments, asdetailed herein, the third substituted aldehyde precursor is anN-protected α-amino aldehyde precursor. In certain other embodiments,the third aldehyde precursor is an alternate substituted aldehydeprecursor, as defined generally. For example, the term “substitutedaldehyde precursor”, as used herein, refers generally to structures ofthe formula R₉(CH₂)_(m)CHO, wherein m is 0-5 and R₉ is NR_(L)R_(M),—OR_(L), —SR_(L), —C(═O)R_(L), —C(═)R_(L), —S(O)₂R_(L), or an aliphatic,heteroaliphatic, aryl, heteroaryl, (aliphatic)aryl,(aliphatic)heteroaryl, (heteroaliphatic)aryl, or(heteroaliphatic)heteroaryl moiety, wherein each occurrence of R_(L) andR_(M) is independently hydrogen, —(C═O)R_(N), —NHR_(N), —(SO₂)R_(N),—OR_(N), or an aliphatic, heteroaliphatic, aryl, or heteroaryl moiety,or R_(L) and R_(M), when taken together form an aryl, heteroaryl,cycloaliphatic, or cycloheteroaliphatic moiety, wherein each occurrenceof R_(N) is independently hydrogen, —OR_(P), —SR_(P), —NHR_(P),—(C═O)R_(P), or an aliphatic, heteroaliphatic, aryl, or heteroarylmoiety, wherein each occurrence of R_(P) is independently hydrogen, aprotecting group, or an aliphatic, heteroaliphatic, aryl, heteroaryl,acyl, alkoxy, aryloxy, alkylthio, arylthio, heteroaryloxy, orheteroarylthio moiety. It will be appreciated that N-protected a-aminoaldehyde precursors are encompassed by the generic term “substitutedaldehyde precursors”. Certain exemplary embodiments are discussed inmore detail below and in the exemplification herein.

As described in more detail in the Exemplification section herein, themethodology described generally above takes advantage of the ability togenerate a series of imine structures and subsequently subject theseimines to suitable reaction conditions to effect cyclization reactions,thus generating the desired alkaloid structures. Thus, in one embodimentof special interest, a first N-protected α-amino aldehyde precursor (XI)is provided, and reacted with a second C-protected α-amino aldehydeprecursor (XI) under suitable conditions to generate atetrahydroisoquinoline core structure (XIII), which reaction comprisesfirst reacting said precursors to effect Schiff-base formation andPictet-Spengler cyclization and generate a tetrahydroisoquinoline coreof structure (XIII); and optionally reacting the tetrahydroisoquinolinecore structure (MII) to further functionalize the core structure at R₃;and wherein the step of reacting said third aldehyde precursor with saidtetrahydrisoquinoline core structure comprises reacting under suitableconditions to effect another Pictet-Spengler cyclization to generate atrimer of aldehydes (XIV) and reacting said trimer (XIV) under suitableconditions to effect cyclization and generate an alkaloid structure(XV).

In certain embodiments of special interest, said first N-protectedα-amino aldehyde precursor, and said second C-protected a-amino aldehydeprecursor have the general structures:

wherein said tetrohydroisoquinoline core has the structure (XIII):

wherein said third aldehyde precursor is R₉(CH₂)_(m)CHO;

wherein said trimer of amino aldehydes has the structure (XIV):

wherein X₁—X₄, R₂—R₈ and m are as defined generally and in subclassesherein;

P₁ is hydrogen or a nitrogen protecting group;

X₅ and X₆ taken together represent a carbon protecting group, optionallysubstituted with

a solid support unit; and

R₉ is NR_(L)R_(M), —OR_(L), —SR_(L), —C(═O)R_(L), —C(═)R_(L),—S(O)₂R_(L), or an aliphatic, heteroaliphatic, aryl, heteroaryl,(aliphatic)aryl, (aliphatic)heteroaryl, (heteroaliphatic)aryl, or(heteroaliphatic)heteroaryl moiety, wherein each occurrence of R_(L) andR_(M) is independently hydrogen, —(C═O)R_(N), —NHR_(N), —(SO₂)R_(N),—OR_(N), or an aliphatic, heteroaliphatic, aryl, or heteroaryl moiety,or R_(L) and R_(M), when taken together form an aryl, heteroaryl,cycloaliphatic, or cycloheteroaliphatic moiety, wherein each occurrenceof R_(N) is independently hydrogen, —OR_(P), —SR_(P), —NHR_(P),—(C═O)R_(P), or an aliphatic, heteroaliphatic, aryl, or heteroarylmoiety, wherein each occurrence of R_(P) is independently hydrogen, aprotecting group, or an aliphatic, heteroaliphatic, aryl, heteroaryl,acyl, alkoxy, aryloxy, alkylthio, arylthio, heteroaryloxy, orheteroarylthio moiety.

It will be appreciated that, in addition to the synthesis of compoundsas described herein, the novel method can be utilized to generate avariety of compounds. For example, the present invention alsocontemplates the synthesis of ecteinascidin analogues where X₁ and R₇taken together are a heterocyclic moiety.

In certain embodiments of special interest herein for the intermediates(XIV) and (XV) R₉ is —NHP₂, wherein P₂ is a nitrogen protecting group,and thus the intermediates have the structures (XIVa) and (XVa):

As mentioned above, in certain other embodiments of the invention,compounds of formula (XV) and (XVa) can be further modified to generatecompounds of general formula (I) including classes and subclassesthereof, as described in more detail herein.

In certain other embodiments for methods as described herein, the thirdsubstituted aldehyde precursor, R₉(CH₂)_(m)CHO is(aliphatic)(C═O)(CH₂)_(m)CHO, (heteroaliphatic)(C═O) (CH₂)_(m)CHO,(aliphatic)(CH₂)_(m)CHO, (heteroaliphatic)(CH₂)_(m)CHO,aryl(aliphatic)(CH₂)_(m)CHO, aryl(heteroaliphatic)(CH₂)_(m)CHO,-heteroaryl(aliphatic)(CH₂)_(m)CHO, orheteroaryl(heteroaliphatic)(CH₂)_(m)CHO, wherein each of the aliphatic,heteroaliphatic, aryl, and heteroaryl moieties is independentlysubstituted or unsubstituted. In certain embodiments, any one or more ofthe aliphatic, heteroaliphatic, aryl or heteroaryl moieties issubstituted with one or more of substituted or unsubstituted amino,substituted or unsubstituted thiol, or substituted or unsubstitutedhydroxyl. In certain exemplary embodiments, as described in more detailin the exemplification herein, the third substituted aldehyde precursoris CH₃(CH₂)₁₋₆CHO; (protecting group)O(CH₂)₁₋₆CHO; (protectinggroup)NH(CH₂)₁₋₆CHO; (protecting group)S(CH₂)₁₋₆CHO; (alkyl)O(C═O)CHO;(aryl)(alkenyl)CHO; heteroaryl)(alkenyl)CHO; (aryl)CHO; orheteroaryl)CHO, wherein any one or more of the aryl, heteroaryl, alkenylor alkyl moieties is substituted or unsubstituted.

In still other exemplary embodiments as described herein X₅ is CN and X₆is a heterocyclic moiety optionally substituted with a solid supportunit.

Thus, the present invention additionally provides a method for thesynthesis of compounds of structure (1) as described above and inclasses and subclasses herein, which method comprises:

(a) providing a compound of formula (XV)

(b) reacting said compound of formula (XV) under sutiable conditions togenerate a compound of formula (I):

wherein X₁—X₄, R₁—R₈, and m are as described above and in classes andsubclasses herein, and

wherein the step of providing a compound of formula (XV) furthercomprises:

-   -   (1) reacting a first N-protected and a second C-protected        x-amino aldehyde precursor having the structures:    -   under suitable conditions to generate a tetrahydroisoquinoline        core having the structure (IX):    -   (2) optionally reacting said tetrahydroisoquinoline core under        suitable conditions to diversify R₃;    -   (3) reacting a third aldehyde precursor having the structure:        R₉(CH₂)_(m)CHO, with said tetrahydroisoquinoline core        structure (XXV) under suitable conditions to generate a trimer        of aldehydes having the structure:    -   (4) reacting said trimer of aldehydes under suitable conditions        to generate a compound of structure (XV).

In certain embodiments of special interest herein for the intermediates(XIV) and (XV) R₉ is —NHP₂, wherein P₂ is a nitrogen protecting group,and thus the intermediates have the structures (XIVa) and (XVa):

As described above, in certain embodiments, the alkaloid structuresgenerated by the method of the present invention represent naturalproduct precursors. Thus, subsequent reaction of these precursorsenables the production of the desired natural products, and, in certainembodiments, the methods of the present invention enables thestereoselective production of the desired natural product precursors andnatural products. In certain embodiments of special interest, themethods as described herein are utilized for the synthesis of analkaloid structure, wherein the alkaloid structure (1) generated is thatof saframycin A or derivatives thereof. In certain other particularlypreferred embodiments, the method is stereoselective and the alkaloidstructure (1) generated is that of -(−) saframycin A and derivativesthereof.

It will be appreciated, however, that the method of the presentinvention can be utilized for the synthesis of any of the compounds asdescribed herein and for classes and subclasses thereof. Additionally,the method of the present invention can be utilized for the synthesis ofnaturally occurring saframycins and related compounds (e.g., forecteinascidins and analogues thereof, where R₇ and X₁ taken together area heterocyclic moiety (methylenedioxy, in certain embodiments)).

It will be appreciated that a variety of experimental conditions can beutilized to effect the synthetic transformations as described generallyabove. The Exemplification sections describe certain experimentalconditions in more detail to enable the production of the compounds ofthe present invention. It will be appreciated however, that althoughcertain reagents are specifically described in the experimentals, avariety of equivalent reagents can also be utilized. In but one example,although LiBr was utilized to effect activation of the iminefunctionality to initiate Pictet-Spengler cyclization, it will beappreciated that a variety of other suitable Lewis acids known in theart may be utilized for the activation of the imine functionality asdescribed in the methodology herein. In certain preferred embodiments,those Lewis acids that will not ionise the amino nitrile are utilized inthe method of the present invention. Additionally, as described herein,the compounds of the present invention can be diversified at a varietyof functional sites (e.g., R₁, R₂, R₃, R₉, oxidation of one or morearomatic rings to quinone moieties, to name a few) either aftersynthesis of the core structure (XV) or during the synthesis of thecompounds. As shown in the Exemplification herein, compounds where R₉ is—NHP₁ can be diversified to generate a variety of analogues asdescribed. Additionally, use of a variety of third aldehyde precursorsin the method of the present invention yields analogues in which thecore ring structure is altered (e.g., —C—C linkages, —C—N linkages,—C—C(═O) linkages etc.). Additionally, as shown in FIGS. 14A and 14B,and as described herein, the N-alkylation reaction can be varied toyield exemplary analogues diversified at R₃.

It will be appreciated that the general synthetic method described abovecan be utilized with solid support techniques. Thus, in certainembodiments of the invention, compounds of the invention are preparedusing a solid support. As described herein, the desired alkaloidprecursors may be modified or reacted directly to effect attachment tothe solid support. The use of a solid support bound component enablesthe use of more rapid combinatorial (parallel or split-pool) techniquesto generate large numbers of compounds more easily. In general, asdescribed in the Exemplification herein, the carbon protected aldehydecan be modified by attachment of a linker moiety (generally an aliphaticor heteroaliphatic moiety) to facilitate attachment of the solidsupport. The solid support bound carbon protected aldehyde is thenreacted under suitable conditions with the first a-amino aldehydedescribed generally above to generate a solid support boundtetrahydroisoquinoline derivative. Finally, reaction under suitableconditions with a third aldehyde precursor results in the cleavage ofthe solid support unit and generation of the desired compound.

A solid support, for the purposes of this invention, is defined as aninsoluble material to which compounds are attached during a synthesissequence. The use of a solid support is advantageous for the synthesisof libraries because the isolation of support-bound reaction productscan be accomplished simply by washing away reagents from thesupport-bound material and therefore the reaction can be driven tocompletion by the use of excess reagents.

Additionally, the use of a solid support also enables the use ofspecific encoding techniques to “track” the identity of the inventivecompounds in the library. A solid support can be any material which isan insoluble matrix and can have a rigid or semi-rigid surface.Exemplary solid supports include, but are not limited to, pellets,disks, capillaries, hollow fibers, needles, pins, solid fibers,cellulose beads, pore-glass beads, silica gels, polystyrene beadsoptionally cross-linked with divinylbenzene, grafted co-poly beads,poly-acrylamide beads, latex beads, dimethylacrylamide beads optionallycrosslinked with N-N′-bis-acryloylethylenediamine, and glass particlescoated with a hydrophobic polymer. One of ordinary skill in the art willrealize that the choice of particular solid support will be limited bythe compatability of the support with the reaction chemistry beingutilized.

It will be appreciated that specific compounds may be attached directlyto the solid support or may be attached to the solid support through alinking reagent. Direct attachment to the solid support may be useful ifit is desired not to detach the library member from the solid support.For example, for direct on-bead analysis of biological/pharmacologicalactivitiy or analysis of the compound structure, a stronger interactionbetween the library member and the solid support may be desirable.Alternatively, the use of a linking reagent may be useful if more facilecleavage of the inventive library members from the solid support isdesired.

Furthermore, any linking reagent used in the present invention maycomprise a single linking molecule, or alternatively may comprise alinking molecule and one or more spacer molecules. A spacer molecule isparticularly useful when the particular reaction conditions require thatthe linking molecule be separated from the library member, or ifadditional distance between the solid support/linking unit and thelibrary member is desired.

Thus, in certain embodiments, libraries of inventive alkaloids can beprepared using established combinatorial methods for solution phase,solid phase, or a combination of solution phase and solid phasesynthesis techniques. The synthesis of combinatorial libraries is wellknown in the art and has been reviewed (see, e.g., “CombinatorialChemistry”, Chemical and Engineering News, Feb. 24, 1997, p. 43;Thompson, L. A., Ellman, J. A., Chem. Rev. 1996, 96, 555.) I0n certainembodiments, the use of solid phase techniques may be desired and thusencoding techniques may also be employed. Specific encoding techniqueshave been reviewed by Czarnik. (Czarnik, A. W., Current Opinion inChemical Biology, 1997, 1, 60.) One of ordinary skill in the art willrealize that the choice of method will depend upon the specific numberof compounds to be synthesized, the specific reaction chemistry, and theavailability of specific instrumentation, such as roboticinstrumentation for the preparation and analysis of libraries. Inparticularly preferred embodiments, the reactions to be performed on thecompound precursors are selected for their ability to proceed in highyield, and in a stereoselective fashion, if desired.

As described in the Exemplification herein, and as shown in FIG. 10, inone embodiment of particular interest, the inventive compounds arepreparing using a modular solid-supported synthesis. As describedgenerally above, the C-protected aldehyde precursor can be furthermodified with a solid support unit. As used herein, the term “solidsupport unit” includes a solid support, as defined herein, andadditionally optionally includes a linker moiety which may be analiphatic, heteroaliphatic, aryl or heteroaryl moiety that facilitatesattachment of the solid support to the intermediate of interest. Incertain embodiments, herein, a solid support unit is attached to thecarbon protecting group (by preparation of a modified group, e.g.,preparation of a siloxymorpholine moiety as described in theExemplification section herein) through a linker unit. It will beappreciated that a variety of linkages and solid supports can beutilized in the method of the invention.

5) Uses, Formulation and Administration

Pharmaceutical Compositions

As discussed above, the present invention provides novel compoundshaving antitumor and antiproliferative activity, and thus the inventivecompounds are useful for the treatment of cancer. Accordingly, inanother aspect of the present invention, pharmaceutical compositions areprovided, wherein these compositions comprise any one of the compoundsas described herein, and optionally comprise a pharmaceuticallyacceptable carrier. In certain embodiments, these compositionsoptionally further comprise one or more additional therapeutic agents.In certain other embodiments, the additional therapeutic agent is ananticancer agent, as discussed in more detail herein.

It will also be appreciated that certain of the compounds of presentinvention can exist in free form for treatment, or where appropriate, asa pharmaceutically acceptable derivative thereof. According to thepresent invention, a pharmaceutically acceptable derivative includes,but is not limited to, pharmaceutically acceptable salts, esters, saltsof such esters, or any other adduct or derivative which uponadministration to a patient in need is capable of providing, directly orindirectly, a compound as otherwise described herein, or a metabolite orresidue thereof, e.g., a prodrug.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts which are, within the scope of sound medical judgement,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well known in the art. For example, S. M. Berge, etal. describe pharmaceutically acceptable salts in detail in J.Pharmaceutical Sciences, 66: 1-19 (1977), incorporated herein byreference. The salts can be prepared in situ during the final isolationand purification of the compounds of the invention, or separately byreacting the free base function with a suitable organic acid. Examplesof pharmaceutically acceptable, nontoxic acid addition salts are saltsof an amino group formed with inorganic acids such as hydrochloric acid,hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid orwith organic acids such as acetic acid, oxalic acid, maleic acid,tartaric acid, citric acid, succinic acid or malonic acid or by usingother methods used in the art such as ion exchange. Otherpharmaceutically acceptable salts include adipate, alginate, ascorbate,aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate,camphorate, camphorsulfonate, citrate, cyclopentanepropionate,digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,glucoheptonate, glycerophosphate, gluconate, hernisulfate, heptanoate,hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate,lactate, laurate, lauryl sulfate, malate, maleate, malonate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate,oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate,phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate,tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts,and the like. Representative alkali or alkaline earth metal saltsinclude sodium, lithium, potassium, calcium, magnesium, and the like.Further pharmaceutically acceptable salts include, when appropriate,nontoxic ammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, loweralkyl sulfonate and aryl sulfonate.

Additionally, as used herein, the term “pharmaceutically acceptableester” refers to esters which hydrolyze in vivo and include those thatbreak down readily in the human body to leave the parent compound or asalt thereof. Suitable ester groups include, for example, those derivedfrom pharmaceutically acceptable aliphatic carboxylic acids,particularly alkanoic, alkenoic, cycloalkanoic and alkanedioic acids, inwhich each alkyl or alkenyl moiety advantageously has not more than 6carbon atoms. Examples of particular esters include formates, acetates,propionates, butyrates, acrylates and ethylsuccinates.

Furthermore, the term “pharmaceutically acceptable prodrugs” as usedherein refers to those prodrugs of the compounds of the presentinvention which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswith undue toxicity, irritation, allergic response, and the like,commensurate with a reasonable benefit/risk ratio, and effective fortheir intended use, as well as the zwitterionic forms, where possible,of the compounds of the invention. The term “prodrug” refers tocompounds that are rapidly transformed in vivo to yield the parentcompound of the above formula, for example by hydrolysis in blood. Athorough discussion is provided in T. Higuchi and V. Stella, Pro-drugsas Novel Delivery Systems, Vol. 14 of the A.C.S. Symposium Series, andin Edward B. Roche, ed., Bioreversible Carriers in Drug Design, AmericanPharmaceutical Association and Pergamon Press, 1987, both of which areincorporated herein by reference.

As described above, the pharmaceutical compositions of the presentinvention additionally comprise a pharmaceutically acceptable carrier,which, as used herein, includes any and all solvents, diluents, or otherliquid vehicle, dispersion or suspension aids, surface active agents,isotonic agents, thickening or emulsifying agents, preservatives, solidbinders, lubricants and the like, as suited to the particular dosageform desired. Remington's Pharmaceutical Sciences, Sixteenth Edition, E.W. Martin (Mack Publishing Co., Easton, Pa., 1980) discloses variouscarriers used in formulating pharmaceutical compositions and knowntechniques for the preparation thereof. Except insofar as anyconventional carrier medium is incompatible with the anti-cancercompounds of the invention, such as by producing any undesirablebiological effect or otherwise interacting in a deleterious manner withany other component(s) of the pharmaceutical composition, its use iscontemplated to be within the scope of this invention. Some examples ofmaterials which can serve as pharmaceutically acceptable carriersinclude, but are not limited to, sugars such as lactose, glucose andsucrose; starches such as corn starch and potato starch; cellulose andits derivatives such as sodium carboxymethyl cellulose, ethyl celluloseand cellulose acetate; powdered tragacanth; malt; gelatin; talc;excipients such as cocoa butter and suppository waxes; oils such aspeanut oil, cottonseed oil; safflower oil; sesame oil; olive oil; cornoil and soybean oil; glycols; such a propylene glycol; esters such asethyl oleate and ethyl laurate; agar; buffering agents such as magnesiumhydroxide and aluminum hydroxide; alginic acid; pyrogen-free water,isotonic saline; Ringer's solution; ethyl alcohol, and phosphate buffersolutions, as well as other non-toxic compatible lubricants such assodium lauryl sulfate and magnesium stearate, as well as coloringagents, releasing agents, coating agents, sweetening, flavoring andperfuming agents, preservatives and antioxidants can also be present inthe composition, according to the judgment of the formulator.

Uses of Compounds and Pharmaceutical Compositions

In yet another aspect, according to the methods of treatment of thepresent invention, tumor cells are killed, or their growth is inhibitedby contacting said tumor cells with an inventive compound orcomposition, as described herein. Thus, in still another aspect of theinvention, a method for the treatment of cancer is provided comprisingadministering a therapeutically effective amount of an inventivecompound, or a pharmaceutical composition comprising an inventivecompound to a subject in need thereof, in such amounts and for such timeas is necessary to achieve the desired result. In certain embodiments ofthe present invention a “therapeutically effective amount” of theinventive compound or pharmaceutical composition is that amounteffective for killing or inhibiting the growth of tumor cells. Thecompounds and compositions, according to the method of the presentinvention, may be administered using any amount and any route ofadministration effective for killing or inhibiting the growth of tumorcells. Thus, the expression “amount effective to kill or inhibit thegrowth of tumor cells”, as used herein, refers to a sufficient amount ofagent to kill or inhibit the growth of tumor cells. The exact amountrequired will vary from subject to subject, depending on the species,age, and general condition of the subject, the severity of theinfection, the particular anticancer agent, its mode of administration,and the like. The anticancer compounds of the invention are preferablyformulated in dosage unit form for ease of administration and uniformityof dosage. The expression “dosage unit form” as used herein refers to aphysically discrete unit of anticancer agent appropriate for the patientto be treated. It will be understood, however, that the total dailyusage of the compounds and compositions of the present invention will bedecided by the attending physician within the scope of sound medicaljudgment. The specific therapeutically effective dose level for anyparticular patient or organism will depend upon a variety of factorsincluding the disorder being treated and the severity of the disorder;the activity of the specific compound employed; the specific compositionemployed; the age, body weight, general health, sex and diet of thepatient; the time of administration, route of administration, and rateof excretion of the specific compound employed; the duration of thetreatment; drugs used in combination or coincidental with the specificcompound employed; and like factors well known in the medical arts.

Furthermore, after formulation with an appropriate pharmaceuticallyacceptable carrier in a desired dosage, the pharmaceutical compositionsof this invention can be administered to humans and other animalsorally, rectally, parenterally, intracisternally, intravaginally,intraperitoneally, topically (as by powders, ointments, or drops),bucally, as an oral or nasal spray, or the like, depending on theseverity of the infection being treated. In certain embodiments, thecompounds of the invention may be administered orally or parenterally atdosage levels of about 0.01 mg/kg to about 50 mg/kg and preferably fromabout 1 mg/kg to about 25 mg/kg, of subject body weight per day, one ormore times a day, to obtain the desired therapeutic effect.

Liquid dosage forms for oral administration include, but are not limitedto, pharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to the active compounds,the liquid dosage forms may contain inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformamide, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor, and sesame oils),glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fattyacid esters of sorbitan, and mixtures thereof. Besides inert diluents,the oral compositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a drug, it is often desirable to slowthe absorption of the drug from subcutaneous or intramuscular injection.This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material with poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolutionwhich, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle. Injectable depot forms are made by forming microencapsulematrices of the drug in biodegradable polymers such aspolylactide-polyglycolide. Depending upon the ratio of drug to polymerand the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions which are compatible with body tissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like. The solid dosage forms of tablets, dragees, capsules, pills,and granules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifing agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions which can beused include polymeric substances and waxes. Solid compositions of asimilar type may also be employed as fillers in soft and hard-filledgelatin capsules using such excipients as lactose or milk sugar as wellas high molecular weight polethylene glycols and the like.

The active compounds can also be in micro-encapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active compound may be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms may alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents. They may optionally contain opacifying agents and canalso be of a composition that they release the active ingredient(s)only, or preferentially, in a certain part of the intestinal tract,optionally, in a delayed manner. Examples of embedding compositionswhich can be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, ear drops, and eye drops are also contemplatedas being within the scope of this invention. Additionally, the presentinvention contemplates the use of transdermal patches, which have theadded advantage of providing controlled delivery of a compound to thebody. Such dosage forms can be made by dissolving or dispensing thecompound in the proper medium. Absorption enhancers can also be used toincrease the flux of the compound across the skin. The rate can becontrolled by either providing a rate controlling membrane or bydispersing the compound in a polymer matrix or gel.

As discussed above, the compounds of the present invention are useful asanticancer agents, and thus may be useful in the treatment of cancer, byeffecting tumor cell death or inhibiting the growth of tumor cells. Ingeneral, the inventive anticancer agents are useful in the treatment ofcancers and other proliferative disorders, including, but not limited tobreast cancer, cervical cancer, colon and rectal cancer, leukemia, lungcancer, melanoma, multiple myeloma, non-Hodgkin's lymphoma, ovariancancer, pancreatic cancer, prostate cancer, and gastric cancer, to namea few. In certain embodiments, the inventive anticancer agents areactive against leukemia cells and melanoma cells, and thus are usefulfor the treatment of leukemias (e.g., myeloid, lymphocytic, myelocyticand lymphoblastic leukemias) and malignant melanomas. In still otherembodiments, the inventive anticancer agents are active against solidtumors and also kill and/or inhibit the growth of multidrug resistantcells (MDR cells).

It will also be appreciated that the compounds and pharmaceuticalcompositions of the present invention can be employed in combinationtherapies, that is, the compounds and pharmaceutical compositions can beadministered concurrently with, prior to, or subsequent to, one or moreother desired therapeutics or medical procedures. The particularcombination of therapies (therapeutics or procedures) to employ in acombination regimen will take into account compatibility of the desiredtherapeutics and/or procedures and the desired therapeutic effect to beachieved. It will also be appreciated that the therapies employed mayachieve a desired effect for the same disorder (for example, aninventive compound may be administered concurrently with anotheranticancer agent), or they may achieve different effects (e.g., controlof any adverse effects).

For example, other therapies or anticancer agents that may be used incombination with the inventive anticancer agents of the presentinvention include surgery, radiotherapy (in but a few examples,y-radiation, neutron beam radiotherapy, electron beam radiotherapy,proton therapy, brachytherapy, and systemic radioactive isotopes, toname a few), endocrine therapy, biologic response modifiers(interferons, interleukins, and tumor necrosis factor (TNF) to name afew), hyperthermia and cryotherapy, agents to attenuate any adverseeffects (e.g., antiemetics), and other approved chemotherapeutic drugs,including, but not limited to, alkylating drugs (mechlorethamine,chlorambucil, Cyclophosphamide, Melphalan, Ifosfamide), antimetabolites(Methotrexate), purine antagonists and pyrimidine antagonists(6-Mercaptopurine, 5-Fluorouracil, Cytarabile, Gemcitabine), spindlepoisons (Vinblastine, Vincristine, Vinorelbine, Paclitaxel),podophyllotoxins (Etoposide, Irinotecan, Topotecan), antibiotics(Doxorubicin, Bleomycin, Mitomycin), nitrosoureas (Carmustine,Lomustine), inorganic ions (Cisplatin, Carboplatin), enzymes(Asparaginase), and hormones (Tamoxifen, Leuprolide, Flutamide, andMegestrol), to name a few. For a more comprehensive discussion ofupdated cancer therapies see, http://www.nci.nih.gov/, a list of the FDAapproved oncology drugs athttp://www.fda.gov/cder/cancer/druglistframe.htm, and The Merck Manual,Seventeenth Ed. 1999, the entire contents of which are herebyincorporated by reference.

In still another aspect, the present invention also provides apharmaceutical pack or kit comprising one or more containers filled withone or more of the ingredients of the pharmaceutical compositions of theinvention, and in certain embodiments, includes an additional approvedtherapeutic agent for use as a combination therapy. Optionallyassociated with such container(s) can be a notice in the form prescribedby a governmental agency regulating the manufacture, use or sale ofpharmaceutical products, which notice reflects approval by the agency ofmanufacture, use or sale for human administration.

Equivalents

The representative examples which follow are intended to help illustratethe invention, and are not intended to, nor should they be construed to,limit the scope of the invention. Indeed, various modifications of theinvention and many further embodiments thereof, in addition to thoseshown and described herein, will become apparent to those skilled in theart from the full contents of this document, including the exampleswhich follow and the references to the scientific and patent literaturecited herein. It should further be appreciated that the contents ofthose cited references are incorporated herein by reference to helpillustrate the state of the art. The following examples containimportant additional information, exemplification and guidance which canbe adapted to the practice of this invention in its various embodimentsand the equivalents thereof.

Exemplification

I Synthesis of Alkaloids:

A. As discussed above, in one aspect of the invention, novel syntheticmethodology for the preparation of alkaloids is provided. In but oneembodiment, this methodology involves the directed condensation ofa-amino aldehyde precursors. As described in more detail below, thesynthesis of saframycin A and intermediates thereof is described.

i) General Description:

Referring to FIGS. 1 and 2, and Scheme 1, a short and enantioselectivesynthetic route to the potent antitumor agent (−)-saframycin A (1), abisquinone alkaloid of microbial origin is provided (for reviews, see,(a) Arai, et al. In The Alkaloids; Brossi, A., Ed.; Academic Press: NewYork, 1983; Vol. 21, Chapter 3 . (b) Remers, W. A. In The Chemistry ofAntitumor Antibiotics; Wiley-Interscience: New York, 1988; Vol. 2,Chapter 3). The route employs a new and powerful synthetic strategyinvolving the directed condensation of optically active α-aminoaldehydes. This strategy evolved from retrosynthetic analysis of 1, asshown, where a series of transformations initiated by the condensationof an aldehyde with an amine (e.g., reductive amination,Pictet-Spengler, and Strecker reactions), was envisioned to assemble thetarget 1 from five simple components: hydrogen cyanide, formaldehyde,and three α-amino aldehydes, two of which (structure 3) are thesame-hence the latent symmetry of 1. The complexity of the analysisarises in the determination of the precise order and stereochemistry ofbonding events that ultimately links the precursors (seven bonds must beformed), and upon consideration of the fundamental issues of stability,reactivity, and protection strategies surrounding the proposed use ofoptically active α-amino aldehydes as synthetic intermediates. Recently,a series of “C-protected” optically active α-amino aldehydes werereported that incorporate an amino nitrile group as a masked aldehyde(Myers et al. J. Am. Chem. Soc. 1999, 121, 8401). Morpholino nitrilederivatives, exemplified by structure 5, were found to be particularlyuseful synthetic intermediates, undergoing condensation reactions withoptically active N-protected α-amino aldehydes with little to noepimerization of either component, thus establishing the basis for thedirected assembly of (−)-saframycin A detailed herein.

Compounds 4 and 5, N- and C-protected versions of the same chiralα-amino aldehyde (3), were prepared in high enantiomeric excess from thesame product of asymmetric alkylation of (−)-pseudoephedrineglycinaride, as previously described (Myers et al. J. Am. Chem. Soc.1999, 121, 8401; Myers et al J. Org. Chem. 1999, 64, 3322). Addition ofN-protected α-amino aldehyde 4 (96% ee, 1.05 equiv) to C-protectedα-amino aldehyde 5 (92% ee, 1 equiv) in dichloromethane at 23° C. in thepresence of sodium sulfate cleanly provided the imine 6 (presumed trans)without detectable epimerization of either α-stereocenter (¹H NMRanalysis, >90% yield, dr ˜95:5). Addition of a saturated solution ofanhydrous lithium bromide in dimethoxyethane to the imine intermediateand warming to 35° C. brought about Pictet-Spengler cyclization toprovide a ˜5:1 mixture of cis and trans tetrahydroisoquinolines,respectively. Flash column chromatography afforded the desired cisproduct (7) in 65-72% yield and 99% ee. The optical purity of 7 wasassayed by HPLC analysis (Chiralcel OD) of the correspondingbis(benzoyl) derivative against an authentic sample of its enantiomer,derived from (+)-pseudoephedrine via ent-4 and ent-5. Lithium ion provedto be optimal for mild and selective Lewis-acid activation of the iminefunction without reaction of the morpholino nitrile. The cis-transselectivity of the cyclization reaction varied markedly as a function ofsolvent and activating agent; for example, use of lithium perchlorate indiethyl ether provided the trans product exclusively. It is alsonoteworthy that the transformation of 6 to 7 is the only step in thesynthetic route that was conducted above ambient temperature.

Introduction of the N-methyl group at this stage of the synthesis wasfound to be optimal. Stirring 7 at 23° C. in the presence of formalin(2.0 equiv) and sodium triacetoxyborohydride (1.5 equiv) in acetonitrileprovided the corresponding N-methylated compound in 94% yield; themorpholino nitrile function was unaffected by the reductive conditions.The N-Fmoc and OTBS protective groups were then cleaved. While thesedeprotections could be performed simultaneously by the action offluoride or hydroxide, sequential removal of the silyl ether with aceticacid-buffered tetrabutylammonium fluoride (2.4 and 1.1 equiv,respectively) followed by cleavage of the carbamate with DBU (1.3 equiv)provided 8 with greater efficiency (92%). Notably, compound 8 showed nopropensity for the primary amine to add to the masked aldehyde undersuch conditions as exposure to silica gel or upon standing in the proticmedium 2,2,2-trifluoroethanol, further highlighting the stability of themorpholino nitrile protective group.

Addition of the third and final α-amino aldehyde component, N-Fmocglycinal (1.5 equiv), to amine 8 (1 equiv) in the presence of sodiumsulfate in deoxygenated dichloromethane at 23° C. produced an imineintermediate which underwent Pictet-Spengler cyclization, also at 23°C., to provide a ≧9:1 mixture of cis and trans tetrahydroisoquinolines,respectively. The desired cis isomer (9) was isolated in 66% yield. Thereaction solvent was again critical for selective formation of thedesired cis tetrahydroisoquinoline; protic solvents affordedpredominantly the trans diastereomer (e.g., in methanol,trans:cis >5:1). With the “trimer” of α-amino aldehydes assembled (9),the C-terminus morpholino nitrile blocking group was then cleaved withanhydrous zinc chloride (Guibe, F. et al. Tetrahedron Lett. 1982, 23,5055) (3.0 equiv) in a mixture of trifluoroethanol and tetrahydrofuran(2:1) at 23° C., producing the key pentacyclic intermediate 2 in 86%yield. This transformation presumably proceeded by the sequentialformation of iminium ion 10, cyclization (addition of the secondaryamine to the iminium ion), expulsion of morpholine, and trapping of theresultant iminium, ion by cyanide. It was necessary to introduceexogenous cyanide (trimethylsilyl cyanide, 2.0 equiv) to ensure completeamino nitrile formation; in the absence of added cyanide, small amounts(5-10%) of the hemiaminal corresponding to hydrolysis of amino nitrile 2were observed, presumably due to adventitious water. Finally, the N-Fmocprotective group of 2 was cleaved with DBU (1.3 equiv) at 23° C. in 88%yield, and the resulting primary amine was acylated with pyruvoylchloride (3.0 equiv) in the presence of N,N-diethylaniline (1.1 equiv)at 0° C. to afford 11 (89%). Oxidative demethylation of thehydroquinones with iodosobenzene (2.5 equiv) in acetonitrile-water (1:1, 0° C.) furnished synthetic (−)-saframycin A in 66% yield (127 mg of(−)-1). The synthetic material was found to be identical in all respects(¹H NMR, ¹³C NMR, IR, HPLC, tlc analysis, and optical rotation) with anauthentic sample of natural saframycin A, kindly provided by ProfessorT. Arai.

In summary, a practical and efficient synthesis of (−)-saframycin A hasbeen developed that proceeds in just 8 steps from the α-amino aldehydeprecursors 4 and 5, in ˜15% overall yield. Significantly, this synthesisillustrates a simple strategy for alkaloid assembly that can be appliedmore generally, and that involves the directed condensation of α-aminoaldehyde precursors in a manner not unlike oligopeptide synthesis (here,with C→N directionality). The present route is suitable for theproduction of 1 in quantity; to date, more than 1 g of 2 and 200 mg of(−)-1 have been prepared. Additionally, this synthetic methodology canalso be utilized for the generation of 2 in significant quantities andthis compound can be further functionalized, as described herein, togenerate the compounds as described herein in suitable quantities fortherapeutic utility.

ii) Experimental Data:

The N- and C-protected α-amino aldehyde derivatives 4 and 5 wereprepared as previously described (Myers et al J. Am. Chem. Soc. 1999,121, 8401) from amide 12, the product of alkylation (Myers et al. J.Org. Chem. 1999, 64, 3322) of (−)-pseudoephedrine glycinamide withbenzylic bromide 13. Bromide 13 was synthesized from2,4-dimethoxy-3-methylphenol (Godfrey, I. M.; Sargent, M. V.; Elix, J.A. J. Chem. Soc., Perkin Trans. 1 1974, 1353-1354) as shown in Scheme 1below.

(a) TBSCl, imidazole, DMF, 23° C., 99%. (b) Br₂, pyridine, DMF, 23° C.,90%. (c) t-BuLi, THF, −90° C.; DMF, −90→23° C.; NaBH₄, EtOH, 0° .C, 77%.(d) PPh₃, Br₂, imidazole, CH₂Cl₂, 0° C. 78%. (e) (−)-pseudoephedrineglycinamide hydrate, LHMDS, LiCl, THF, 0° C., 74%.

Tetrahydroisoguinoline 7

A solution of aldehyde 4 (433 mg, 0.752 mmol, 1.05 equiv) indichloromethane (7.2 mL) was added to a solid mixture of amine 5 (240mg, 0.716 mmol, 1 equiv) and sodium sulfate (2.03 g, 14.3 mmol, 20.0equiv). The resulting suspension was stirred rapidly for 75 min at 23°C., then was filtered through a plug of cotton. The filtrate wasconcentrated and the residue was dried azeotropically by concentrationfrom a 3-mL portion of toluene to afford a white foam. ¹H NMR analysis(CDCl₃) showed two diastereomers of imine 6 to be present in a ratio of˜95:5. Anhydrous lithium bromide (1.62 g, 18.7 mmol, 26 equiv) and1,2-dimethoxyethane (14.3 mL) were then added sequentially to the imineresidue, and the mixture was held in a sonicator for 5 min. Theresultant suspension was warmed to 35° C. and was held at thattemperature for 17.5 h before being cooled to 23° C. The mixture wasdiluted with ethyl acetate (20 mL) and was washed with three 20-mLportions of 4:1 saturated aqueous sodium chloride solution-saturatedaqueous sodium bicarbonate solution. The organic layer was dried oversodium sulfate and was concentrated. ¹H NMR analysis (CDCl₃) of theresidue showed a ˜5:1 mixture of cis:trans tetrahydroisoquinolines to bepresent. The crude product was purified by flash column chromatography(100:1 dichloromethane-methanol) to afford tetrahydroisoquinoline 7 asan off-white solid (460 mg, 72%). HPLC analysis (Chiralcel OD, 4%2-propanol-0.3% diethylamine-hexanes, 0.30 mL/min, 254 nm detection,˜0.1 mg injection) of bis(benzoyl)-7 (PhCOCl, Et₃N, DMAP, CH₂Cl₂, 23°C.) established an enantiomeric excess of 99% (t,7): 79.2 min,tr(ent-7): 89.2 min).

(R)-Morpholino nitrile diastereomer, 7: ¹H NMR (400 MHz, CDCl₃), δ 7.75(d, 2 H, J=7.7 Hz, ArH), 7.51 (d, 1 H, J=8.4 Hz, ArH), 7.44 (d, 1 H,J=7.3 Hz, ArH), 7.40-7.36 (m, 2 H, ArH), 7.30-7.24 (m, 2 H, ArH), 6.35(s, 1 H, ArH), 6.20 (s, 1 H, ArOH), 5.70 (d, 1 H, J=7.0 Hz, NHFmoc),4.82 (br s, 1 H, ArCHN), 4.58 (m, 1 H, CHNHFmoc), 4.43 (dd, 1 H, J=9.7,6.8 Hz, OCH₂CH), 4.19-4.06 (m, 2 H, OCH₂CH), 3.83-3.74 (m, 4 H,CH₂OCH₂), 3.79 (s, 3 H, ArOCH₃), 3.69 (s, 3 H, ArOCH₃), 3.65 (s, 3 H,ArOCH₃), 3.57 (s, 3 H, ArOCH₃), 3.49 (d, 1 H, J=9.9 Hz, CHC≡N), 3.21(dd, 1 H, J=14.6, 2.0 Hz, CH₂ArOH), 3.15-3.08 (m, 1 H, CHCH≡N), 2.87(dd, 1 H, J=13.9, 11.7 Hz, CH₂ArOTBS), 2.73 (m, 2 H, CH₂NCH₂), 2.58 (m,2 H, CH₂NCH₂), 2.34 (dd, 1 H, J=14.4, 11.2 Hz, CH₂ArOH), 2.25 (s, 3 H,ArCH₃), 2.19 (s, 3 H, ArCH₃), 2.15 (dd (obsc), 1 H, CH₂ArOTBS), 0.96 (s,9 H, SiC(CH₃)₃), 0.10 (s, 3 H, SiCH₃), 0.09 (s, 3 H, SiCH₃). ¹³C NMR(100 MHz, CDCl₃), δ 156.4, 151.3, 149.1, 148.6, 145.0, 144.4, 144.2,142.5, 141.3, 127.6, 127.0, 125.1, 125.0, 124.6, 122.4, 120.2, 119.9,119.6, 114.7, 66.8, 66.7, 64.1, 60.7, 60.6, 60.4, 59.8, 56.6, 55.0,50.3, 47.3, 28.9, 28.5, 25.7, 18.2, 9.8, 9.6, 4.6. FTIR (neat film),cm⁻¹ 3345 (w, br, OH/NH), 2934 (s), 2251 (w, C≡N), 1715 (s, C═O), 1471(s). HRMS (ES) Calcd for C₅₀H₆₅N₄O₉Si (M+H)⁺: 893.4521, Found: 893.4535.

(S)-Morpholino nitrile diastereomer, 7: ¹H NMR (400 MHz, CDCl₃), δ 7.74(d, 2 H, J=7.3 Hz, ArH), 7.48 (t, 2 H, J=8.0 Hz, ArH), 7.40-7.35 (m, 2H, ArH), 7.32-7.24 (m, 2 H, ArH), 6.37 (s, 1 H, ArH), 6.29 (s, 1 H,ArOH), 5.78 (d, 1 H, J=7.2 Hz, NHFmoc), 4.72 (br s, ArCHN), 4.37-4.28(m, 1 H (CHNHFmoc), 1 H (OCH₂CH)), 4.17-4.11 (m, 2 H, OCH₂CH), 3.78-3.74(m, 4 H, CH₂OCH₂), 3.76 (s, 3 H, ArOCH₃), 3.67 (s, 6 H, 2×ArOCH₃), 3.63(s, 3 H, ArOCH₃), 3.46 (d, 1 H, J=7.3 Hz, CHCON), 3.24-3.18 (m, 1 H(CH₂ArOH), 1 H (CHCHC□N)), 2.99 (dd, 1 H, J=13.9, 10.6 Hz, CH₂ArOTBS),2.75-2.71 (m, 2 H, CH₂NCH₂), 2.64-2.60 (m, 2 H, CH₂NCH₂), 2.37-2.28 (m,1 H (dd, CH₂ArOTBS), 1 H (dd, CH₂ArOH)), 2.22 (s, 3 H, ArCH₃), 2.19 (s,3 H, ArCH₃), 0.95 (s, 9 H, SiC(CH₃)₃), 0.10 (s, 3 H, SiCH₃), 0.09 (s, 3H, SiCH₃). ¹³C NMR (100 MHz, CDCl₃), δ 156.5, 151.4, 148.7, 148.6,144.9, 144.3, 144.1, 143.9, 142.5, 141.2, 127.6, 127.0, 126.8, 125.2,125.1, 124.2, 122.4, 120.4, 119.9, 116.1, 66.9, 66.6, 65.3, 60.8, 60.6,59.8, 57.1, 54.6, 51.3, 50.9, 47.2, 29.2, 27.8, 25.7, 18.2, 9.8, 9.5,−4.6. FTIR (neat film), cm⁻¹ 3360 (m, br, OH/NH), 2934 (s), 2249 (w,C≡N), 1715 (s, C═O), 1480 (s). HRMS (ES) Calcd for C₅₀H₆₅N₄O₉Si (M+H)⁺:893.452 1, Found: 893.4545.

N-Methyl-7

Formalin (554 μL, 7.39 mmol, 2.0 equiv) and sodium triacetoxyborohydride(1.17 g, 5.52 mmol, 1.5 equiv) were added sequentially to a solution ofamine 7 (3.30 g, 3.69 mmol, 1 equiv) in acetonitrile (25 mL) at 23° C.After 30 min, the cloudy mixture was diluted with ethyl acetate (75 mL)and was washed with two 50-mL portions of 1:1 saturated aqueous sodiumchloride solution-saturated aqueous sodium bicarbonate solution. Theorganic layer was dried over sodium sulfate and was concentrated. Theresidue was purified by flash column chromatography (40% ethylacetate-hexanes) to afford N-methyl-7 as a white solid (3.29 g, 98%).

(R)-Morpholino nitrile diastereomer, N-methyl-7: ¹H NMR (400 MHz,toluene-d₈), ˜10:1 mixture of atropisomers, major signals only, δ 7.50(d, 2 H, J=7.0 Hz, ArE), 7.39 (d, 1 H, J=6.6 Hz, ArH), 7.31 (d, 1 H,J=7.0 Hz, ArH), 7.21-7.13 (m, 4 H, ArE), 7.03 (s, 1 H, ArH), 6.09 (br s,1 H, ArOH), 5.46 (d, 1 H, J=8.8 Hz, NHFmoc), 4.39-4.32 (m, 1 H,CHNHFmoc), 4.25 (d, 1 H, J=8.0 Hz, ArCHN), 4.03-3.98 (m, 1 H, OCH₂CH),3.91-3.84 (m, 2 H, OCH₂CH), 3.58 (s, 3 H, ArOCH₃), 3.54 (s, 3 H,ArOCH₃), 3.46-3.43 (m, 4 H (CH₂OCH₂), 1 H (CHCHC□N)), 3.44 (s, 3 H,ArOCH₃), 3.35 (dd (obsc), 1 H, CH₂ArOTBS), 3.34 (s, 3 H, ArOCH₃),3.16-3.10 (m, 1 H (d, CHC□N), 1 H (dd, CH₂ArOTBS)), 3.00 (dd, 1 H,J=15.0, 12.4 Hz, CH₂ArOH), 2.43-2.35 (m, 2 H (CH₂NCH₂), 1 H (CH₂ArOH)),2.41 (2 s, 3 H each, ArCH₃ and NCH₃), 2.28 (s, 3 H, ArCH₃), 2.24-2.21(m, 2 H, CH₂NCH₂), 1.05 (s, 9 H, SiC(CH₃)₃), 0.26 (s, 3 H, SiCH₃), 0.24(s, 3 H, SiCH₃). ¹³C NMR (100 MHz, CDCl₃), two atropisomers, δ 156.1,151.5, 148.5, 148.3, 144.8, 144.1, 144.0, 143.9, 142.3, 141.1, 127.5,127.0, 126.9, 126.7, 125.2, 125.0, 124.9, 123.5, 122.6, 120.4, 119.8,116.0, 66.7, 66.6, 64.6, 63.5, 61.1, 61.0, 60.6, 59.7, 57.5, 52.0, 47.1,31.1, 25.7, 24.4, 18.1, 9.9, 9.5, −4.6. FTIR (neat film), cm⁻¹ 3357 (m,OH/NH), 2936 (s), 2250 (w, C≡N), 1715 (s, C═O), 1480 (s). HRMS (ES)Calcd for C₅₁H₆₇N₄O₉Si (M+H)⁺: 907.4677, Found: 907.4666.

(S)-Morpholino nitrile diastereomer, N-methyl-7: ¹H NMR (400 MHz,toluene-d₈), ˜7:1 mixture of atropisomers, major signals only, δ 7.49(d, 2 H, J=6.9 Hz, ArH), 7.35 (d, 1 H, J=7.1 Hz, ArH), 7.28 (dd, 1 H,J=6.3, 2.8 Hz, ArH), 7.23-7.18 (m, 4 H, ArH), 6.88 (s, 1 H, ArH), 5.48(s, 1 H, ArOH), 4.90 (d, 1 H, J=9.6 Hz, NHFmoc), 4.24-4.15 (m, 1 H,CHNHFmoc), 4.09 (d, 1 H, J=10.7 Hz, ArCHN), 3.92 (dd, 1 H, J=8.2, 5.5Hz, OCH₂CH), 3.89-3.76 (m, 1 H (CH₂CHNHFmoc), 1 H (CHC≡N), 2 H(OCH₂CH)), 3.69 (s, 3 H, ArOCH₃), 3.55 (s, 3 H, ArOCH₃), 3.50-3.48 (m, 4H, CH₂OCH₂), 3.43 (s, 3 H, ArOCH₃), 3.39 (dd, 1 H, J=15.4, 6.2 Hz,CH₂CHCHC≡N), 3.19 (s, 3 H, ArOCH₃), 2.76-2.67 (m, 1 H (CH₂CHCHC≡N), 1 H(CHCHC≡N)), 2.64 (s, 3 H, NCH₃), 2.61 (dd, 1 H, J=13.7, 9.8 Hz,CH₂CHNHFmoc), 2.42-2.32 (m, 4 H, CH₂NCH₂), 2.27 (s, 3 H, ArCH₃), 2.00(s, 3 H, ArCH₃), 1.06 (s, 9 H, SiC(CH₃)₃), 0.23 (s, 3 H, SiCH₃), 0.22(s, 3 H, SiCH₃). ¹³C NMR (100 MHz, CDCl₃), two atropisomers, δ 155.8,151.6, 148.8, 148.6, 144.6, 144.1, 144.0, 143.7, 142.7, 141.1, 141.0,127.6, 127.5, 127.0, 126.9, 126.6, 125.1, 123.3, 122.3, 121.3, 120.6,119.8, 117.3, 66.8, 66.5, 63.6, 61.2, 61.0, 60.7, 60.0, 59.8, 54.9,50.2, 49.0, 47.1, 33.5, 25.7, 23.6, 18.1, 9.9, 9.5, −4.6. FTIR (neatfilm), cm⁻¹ 3402 (w, br, OH/NH), 2952 (m), 2252 (w, C≡N), 1713 (s, C═O),1480 (s). HRMS (ES) Calcd for C₅₁H₆₇N₄O₉Si (M+H)⁺: 907.4677, Found:907.4713.

Amine 8

Acetic acid (530 μL, 9.28 mmol, 2.4 equiv) and tetrabutylammoniumfluoride (1.0 M in tetrahydrofuran, 4.24 mL, 4.24 mmol, 1.1 equiv) wereadded sequentially to a solution of N-methyl-7 (3.50 g, 3.86 mmol, 1equiv) in tetrahydrofuran (7.7 mL) at 0° C. The solution was then warmedto 23° C. After 1.5 h, the solution was diluted with 25% saturatedaqueous sodium bicarbonate solution (25 mL) and was extracted with ether(3×25 mL). The combined organic layers were dried over sodium sulfateand were concentrated to afford N-Fmoc-8 as a white foam (3.06 g, >99%).A portion of the residue (1.22 g, 1.54 mmmol, 1 equiv) was dissolved indichloromethane (4.4 mL), and the resulting solution was treated with1,8-diazabicyclo[5.4.0]undec-7-ene (DBU, 300 μL, 2.01 mmol, 1.3 equiv)to cleave the N-Fmoc group. After 30 min, the reaction solution wasloaded directly onto a flash chromatography column. Elution with 25:1dichloromethane-methanol afforded amine 8 as a solid (845 mg, 96%).

(R)-Morpholino nitrile diastereomer, 8: ¹H NMR (400 MHz, CDCl₃), δ 6.86(s, 1 H, ArH), 3.84 (s, 3 H, ArOCH₃), 3.81-3.76 (m, 4 H, CH₂OCH₂), 3.76(s, 3 H, ArOCH₃), 3.72 (d(obsc), 1 H, ArCHNCH₃), 3.70 (s, 3 H,ArOCH₃),3.66 (d, 1 H, J=2.7 Hz, CHC≡N), 3.52 (dd, 1 H, J=13.2, 2.2 Hz,CH₂CHNH₂), 3.33 (dd, 1 H, J=14.7, 4.4 Hz, CH₂CHCHC≡N), 2.97-2.91 (m, 1H, CHNH₂), 2.80-2.76 (m, 2 H, CH₂NCH₂), 2.74 (m, 1 H, CHCHC≡N),2.67-2.60 (m, 2 H, CH₂NCH₂), 2.61 (dd (obsc), 1 H, CH₂CHNH₂), 2.60 (s, 3H, NCH₃), 2.40 (dd, 1 H, J=14.7 12.8 Hz, CH₂CHCHC═N), 2.23 (s, 3 H,ArCH₃), 2.21 (s, 3 H, ArCH₃). ¹³C NMR (100 MHz, CDCl₃), δ 150.5, 148.2,146.8, 145.6, 144.4, 143.9, 128.2, 124.5, 124.2, 123.4, 121.8, 116.4,114.3, 67.2, 66.7, 64.8, 62.0, 61.2, 60.7, 60.5, 60.2, 52.2, 46.8, 33.6,23.5, 9.9, 9.5. FTIR (neat film), cm⁻¹ 3364 (m, br, OH/NH), 2938 (s),2250 (w, C≡N), 1455 (s). HRMS (ES) Calcd for C₃₀H₄₃N₄O₇ (M+H)⁺:571.3132, Found: 571.3106.

(S)-Morpholino nitrile diastereomer, 8: ¹H NMR (400 MHz, CDCl₃), δ 6.68(s, 1 H, ArH), 3.85 (s, 3 H, ArOCH₃), 3.84-3.73 (m, 4 H (CH₂OCH₂), 1 H(ArCHNCH₃)), 3.78 (s, 3 H, ArOCH₃), 3.75 (s, 3 H, ArOCH₃), 3.69 (dd, 1H, J=10.3, 3.0 Hz, CH₂CHNH₂), 3.64 (s, 3 H, ArOCH₃), 3.62 (d, 1 H, J=9.5Hz, CHC≡N), 3.31 (dd, 1 H, J=15.6, 6.4 Hz, CH₂CHCHC≡N), 2.97-2.91 (m, 1H, CHNH₂), 2.90-2.83 (m, 1 H, CHCHC≡N), 2.78-2.73 (m, 2 H, CH₂NCH₂),2.64-2.59 (m, 2 H, CH₂NCH₂), 2.59 (s, 3 H, NCH₃), 2.35 (dd, 1 H, J=13.6,10.6 Hz, CH₂CHNH₂), 2.24 (s, 3 H, ArCH₃), 2.22 (s, 3 H, ArCH₃), 2.10(dd, 1 H, J=15.7, 11.6 Hz, CH₂CHCHC≡N). ¹³C NMR (100 MHz, CDCl₃), δ150.4, 148.5, 146.1, 145.3, 144.6, 144.2, 128.4, 124.6, 124.2, 123.3,121.1, 117.1, 114.4, 67.2, 66.8, 66.7, 61.2, 61.1, 60.7, 60.5, 60.4,56.3, 50.5, 49.3, 35.0, 23.7, 9.9, 9.5. FTIR (neat film), cm⁻¹ 3360 (m,br, OH/NH), 2941 (m), 2251 (w, C≡N), 1455 (s). HRMS (ES) Calcd for C₃₀H₄₃N₄O₇ (M+H)⁺: 571.3132, Found: 571.3146.

Tetrahydroisoquinoline 9

A deoxygenated (3 freeze-pump-thaw cycles) solution of N-Fmoc glycinal(190 mg, 0.675 mmol, 1.2 equiv) in dichloromethane (11.3 mL) wastransferred by cannula to a solid mixture of amine 8 (322 mg, 0.564mmol, 1 equiv) and sodium sulfate (1.20 g, 8.45 mmol, 15 equiv). Theresulting suspension was stirred for 17.5 h at 23° C., then was filteredthrough a plug of cotton, rinsing with a 6-mL portion ofdichloromethane. The filtrate was concentrated and the residue waspurified by flash column chromatography (50→70% ethylacetate-hexanes-ethyl acetate→200:1 ethyl acetate-methanol) to affordtetrahydroisoquinoline 9 as a solid (299 mg, 64%).

(R)-Morpholino nitrile diastereomer, 9: ¹H NMR (400 MHz, CDCl₃), signalsbroadened due to atropisomerism, δ 7.76 (dd, 2 H, J=7.3, 3.5 Hz, ArH),7.57 (t, 2 H, J=7.4 Hz, ArH), 7.31 (m, 2 H, ArH), 7.28 (m, 2 H, ArH),5.38 (br s, 1 H, NHFmoc), 4.404.31 (m, 1 H (CHCH₂NHFmoc), 2 H (OCH₂CH)),4.23 (m, 1 H, OCH₂CH), 3.90 (m, 1 H, CH₂NHFmoc), 3.84-3.65 (m, 1 H(CH₃NCHCHNH), 4 H (CH₂OCH₂)), 3.76 (s, 3 H, ArOCH₃), 3.74 (s, 3 H,ArOCH₃), 3.67 (s, 3 H, ArOCH₃), 3.65 (s, 3 H, ArOCH₃), 3.58-3.56 (m, 1 H(CHC≡N), 1 H (ArCH₂CHHH)), 3.43 (m, 1 H, CH₂NHFmoc), 3.28 (dd, 1 H,CH₂CHNCH₃), 2.86 (m, 1 H, CH₂CHNCH₃), 2.75-2.71 (m, 2 H, CH₂NCH₂), 2.68(m, 1 H, ArCH₂CHNH), 2.62 (br s, 3 H, NCH₃), 2.60 (m, 2 H, CH₂NCH₂),2.39 (m, 1 H (CH₂CHNCH₃), 1 H (ArCH₂CHNH)), 2.21 (s, 6 H, 2×ArCH₃). ¹³CNMR (100 MHz, CDCl₃), δ 157.1, 149.1, 148.4, 145.5, 144.0, 143.7, 143.3,141.8, 141.2, 127.6, 127.0, 125.9, 125.1, 123.3, 122.7, 122.3, 120.4,119.9, 116.0, 66.8, 66.6, 65.9, 65.0, 61.1, 60.7, 60.6, 60.5, 58.9,52.4, 52.2, 51.8, 47.2, 46.4, 26.8, 24.0, 9.6, 9.5. FTIR (neat film),cm⁻¹ 3278 (m, OH/NH), 2940 (m), 2249 (w, C≡N), 1705 (s, C═O), 1450 (s).HRMS (ES) Calcd for C₄₇H₅₆N₅O₉ (M+H)⁺: 834.4078, Found: 834.4106.

(S)-Morpholino nitrile diastereomer, 9: ¹H NMR (500 MHz, CDCl₃), signalsbroadened due to atropisomerism, δ 7.76 (d, 2 H, J=7.0 Hz, ArH), 7.57(d, 2 H, J=6.5 Hz, ArH), 7.39 (m, 2 H, ArH), 7.30 (m, 2 H, ArH), 5.44(br s, 1 H, NHFmoc), 4.42 (dd, 1 H, OCH₂CH), 4.35 (m, 1 H CHCH₂NHFmoc),4.28 (t, 1 H, OCH₂CH), 4.22 (dd, 1 H, OCH₂CH), 3.85-3.82 (m, 1 H(CH₂NHFmoc), 1 H (CH₂CHNCH₃)), 3.75-3.65 (m, 4 H (CH₂OCH₂), 1 H(CHC≡N)), 3.74 (br s, 9 H, 3×ArOCH₃), 3.65 (s, 3 H, ArOCH₃), 3.37 (m, 1H, CH₂NHFmoc), 3.32 (dd, 1 H, ArCH₂CHNH), 3.27 (m, 1 H, CH₃NCHCHNH),2.87 (m, 1 H, CH₂CHNCH₃), 2.74 (m, 1 H, ArCH₂CHNH), 2.70 (m, 2 H,CH₂NCH₂), 2.59 (br s, 3 H, NCH₃), 2.53 (m, 2 H, CH₂NCH₂), 2.36 (dd, 1 H,CH₂CHNCH₃), 2.24 (s, 3 H, ArCH₃), 2.22 (s, 3 H, ArCH₃), 2.13 (dd, 1 H,ArCH₂CHHH). ¹³C NMR (125 MHz, CDCl₃), δ 157.0, 149.1, 148.6, 145.3,144.1, 143.9, 143.7, 141.8, 141.2, 127.6, 127.0, 125.8, 125.1, 123.3,122.9, 122.5, 121.9, 120.7, 119.9, 116.3, 66.8, 66.6, 66.1, 62.1, 61.1,60.7, 60.5, 60.3, 57.0, 52.0, 50.5, 48.9, 47.2, 27.7, 24.0, 9.5, 6.0.FTIR (neat film), cm⁻¹ 3274 (m, OH/NH), 2939 (m), 2249 (w, C□N), 1704(s, C═O), 1451 (s). HRMS (ES) Calcd for C₄₇H₅₆N₅O₉ (M+H)⁺: 834.4078,Found: 834.4103.

Pentacyclic Intermediate 2

A solution of anhydrous zinc chloride in tetrahydrofuran (0.50 M, 4.26mL, 2.14 mmol, 3.0 equiv) and trimethylsilyl cyanide (190 μL, 1.42 mmol,2.0 equiv) were added sequentially to a solution of morpholino nitrile 9(595 mg, 0.713 mmol, 1 equiv) in 2,2,2-trifluoroethanol (8.5 mL) at 23°C. After 7 h, an aqueous solution of EDTA (20 mL, 0.20 M(ethylenedinitrilo)tetraacetic acid, disodium salt-0.40 M sodiumhydroxide, pH 10) was added, and the resulting mixture was extractedwith ethyl acetate (2×25 mL). The combined organic layers were washedwith a 20-mL portion of 1:1 saturated aqueous sodium chloridesolution-saturated aqueous sodium bicarbonate solution, then were driedover sodium sulfate and were concentrated. Purification of the residueby radial chromatography (40%→60% ethyl acetate-hexanes) furnishedpentacyclic intermediate 2 as a white solid (464 mg, 87%).

¹H NMR (500 MHz, CDCl₃), ˜5:1 mixture of rotamers, major signals only, δ7.75 (t, 2 H, J=7.9 Hz, ArH), 7.46-7.39 (m, 4 H, ArH), 7.30 (t, 2 H,J=7.4 Hz, ArH), 5.63 (s, 1 H, ArOH), 5.54 (s, 1 H, ArOH), 4.54 (t, 1 H,J=5.8 Hz, NHFmoc), 4.32 (dd, 1 H, J=10.8, 6.8 Hz, OCH₂CH), (dd, 1 H,J=10.8, 6.3 Hz, OCH₂CH), 4.14 (br s, 1 H, ArCHNCH₃), 4.12 (app t, 1 H,J=4.4 Hz, CHCH₂NHFmoc), 4.06 (app t, 1 H, J=6.4 Hz, OCH₂CH), 3.73 (s, 3H, ArOCH₃), 3.68 (br s, 1 H, CHC□N), 3.60 (2 s, 3 H each, 2×ArOCH₃),3.55 (s, 3 H, ArOCH₃), 3.28 (br d, 1 H, J=7.6 Hz, CHCHC≡N), 3.24-3.20(m, 1 H (ArCHCHN), 1 H (ArCHCHCH₂), 1 H (CH₂NHFmoc)), 3.11-3.06 (m, 1 H,CH₂NHFmoc), 2.96 (dd, 1 H, J=18.5, 7.8 Hz, ArCH₂CHNCH₃), 2.33 (d, 1 H,J=18.5 Hz, ArCH₂CHNCH₃), 2.31 (s, 3 H, NCH₃), 2.18 (s, 3 H, ArCH₃), 2.11(s, 3 H, ArCH₃), 1.88 (dd, 1 H, J=15.7, 12.5 Hz, ArCHCHCH₂). ¹³C N (100MHz, CDCl₃), δ 156.0, 148.2, 144.0, 143.7, 143.6, 143.2, 141.6, 141.2,127.6, 127.0, 125.0, 124.8, 123.8, 122.3, 122.1, 119.8, 119.7, 118.5,118.1, 116.8, 65.9, 60.7, 60.6, 59.6, 57.1, 56.6, 56.5, 55.2, 47.2,45.3, 41.7, 25.7, 21.2, 9.6. FTIR (neat film), cm⁻¹ 3395 (m, br, OH/NH),2938 (m), 2250 (w, C≡N), 1714 (s, C═O), 1463 (s). HRMS (ES) Calcd forC₄₃H₄₇N₄O₈ (M+H)⁺: 747.3394, Found: 747.3424.

N-Fmoc Cleavage of Pentacyclic Intermediate 2

1,8-Diazabicyclo[5.4.0]undec-7-ene (DBU, 140 μL, 0.936 mmol, 1.3 equiv)was added to a solution of pentacyclic intermediate 2 (538 mg, 0.720mmol, 1 equiv) in dichloromethane (2.9 mL) at 23° C. After 30 min, thereaction solution was loaded directly onto a flash chromatographycolumn. Elution with 25:1→20:1 dichloromethane-methanol afforded thedeprotected product as a white solid (330 mg, 87%).

¹H NMR (500 MHz, CDCl₃), δ 4.16 (br s, 1 H, ArCHNCH₃), 4.03 (br d, 1 H,J=3.2 Hz, CHCH₂NH₂), 4.00 (d, 1 H, J=2.1 Hz, CHC≡N), 3.74 (2 s, 3 Heach, 2×ArOCH₃), 3.68 (s, 3 H, ArOCH₃), 3.61 (s, 3 H, ArOCH₃), 3.39 (brd, 1 H, J=7.6 Hz, CHCHC≡N), 3.24 (app dd, 1 H, J=11.8, 2.4 Hz,ArCH₂CHCHAr), 3.20 (dd, 1 H, J=15.7, 2.1 Hz, ArCH₂CHCHAr), 3.07 (dd, 1H, J=18.4, 8.0 Hz, CH₂CHCHC≡N), 2.84 (dd, 1 H, J=13.2, 1.8 Hz, CH₂NH₂),2.60 (d, 1 H, J=13.4, 5.9 Hz, CH₂NH₂), 2.43 (d, 1 H, J=18.4 Hz,CH₂CHCHC≡N), 2.32 (s, 3 H, NCH₃), 2.22 (s, 3 H, ArCH₃), 2.18 (s, 3 H,ArCH₃), 1.83 (dd, 1 H, J=15.3, 11.9 Hz, ArCH₂CHCHAr). ¹³C NMR (125 MHz,CDCl₃), δ 148.3, 148.0, 144.6, 143.3, 143.1, 142.6, 124.5, 123.8, 122.5,122.0, 120.1, 118.2, 117.0, 60.7, 60.6, 60.5, 60.4, 59.9, 59.2, 56.5,56.3, 55.1, 46.0, 41.7, 25.8, 21.5, 9.5 (2 C). FTIR (neat film), cm⁻¹3356 (w, br, OH/NH), 2936 (m), 2250 (w, C≡N), 1463 (s). HRMS (ES) Calcdfor C₂₈H₃₇N₄O₆ (M+H)⁺: 525.2713, Found: 525.2690.

Pyruvamide 11

N,N-Diethylaniline (34 μL, 0.21 mmol, 1.1 equiv) and pyruvoyl chloride(65 μL, 0.61 mmol, 3.0 equiv) were added sequentially to a solution ofthe primary amine (107 mg, 0.204 mmol, 1 equiv) in dichloromethane (4.1mL) at 0° C. After 30 min, a 10-mL portion of half-saturated aqueoussodium bicarbonate solution was added, and the resulting mixture wasextracted with ethyl acetate (10 mL). The organic layer was washed onceeach with 10-mL portions of half-saturated aqueous sodium bicarbonatesolution and 1:1 saturated aqueous sodium chloride solution-saturatedaqueous sodium bicarbonate solution, then was dried over sodium sulfateand was concentrated. Flash column chromatography (60% ethylacetatehexanes) afforded pyruvamide 11 as a white solid (105 mg, 87%).

¹H NMR (400 MHz, CDCl₃), δ 6.44 (t, 1 H, J=5.6 Hz, NHCO), 5.71 (s, 1 H,ArOH), 5.59 (s, 1 H, ArOH), 4.23 (br s, 1 H, CHCH₂NHCO), 4.15 (d, 1 H,J=2.1 Hz, ArCHNCH₃), 4.02 (d, 1 H, J=2.4 Hz, CHC≡N), 3.79 (s, 3 H,ArOCH₃), 3.73 (s, 3 H, ArOCH₃), 3.66 (app dt (obsc), 1 H, CH₂NHCO), 3.65(s, 3 H, ArOCH₃), 3.58 (s, 3 H, ArOCH₃), 3.41 (br d, 1 H, J=7.9 Hz,CHCHC≡N), 3.29 (app dt, 1 H, J=13.6, 4.7 Hz, CH₂NHCO), 3.24-3.20 (m, 1 H(ArCHCHCH₂Ar), 1 H (ArCHCHCH₂Ar)), 3.05 (dd, 1 H, J=18.5, 8.1 Hz,CH₂CHCHC≡N), 2.48 (d, 1 H, J=18.5 Hz, CH₂CHCHC≡N), 2.30 (s, 3 H, NCH₃),2.23 (s, 3 H, COCH₃), 2.18 (s, 3 H, ArCH₃), 2.16 (s, 3 H, ArCH₃), 1.92(dd, 1 H, J=16.2, 12.1 Hz, ArCHCHCH₂). ¹³C NMR (125 MHz, CDCl₃), δ196.1, 159.9, 148.4, 148.2, 143.5, 143.3 (2 C), 141.5, 125.0, 123.5,122.6, 122.4, 118.0, 117.5, 116.4, 60.8, 60.7, 60.4, 59.9, 59.6, 56.5 (2C), 56.3, 55.0, 41.8, 41.5, 25.8, 24.3, 21.1, 9.6 (2 C). FTIR (neatfilm), cm⁻¹ 3378 (m, br, OH/NH), 2937 (m), 2251 (w, C≡N), 1720 (m, C═O),1682 (s, C═O), 1463 (s). HRMS (ES) Calcd for C₃₁H₃₉N₄O₈ (M+H)⁺:595.2768, Found: 595.2739.

(−)-Saframycin A (1)

Iodosobenzene (189 mg, 0.859 mmol, 2.5 equiv) was added to a solution ofpyruvamide 11 (203 mg, 0.341 mmol, 1 equiv) in 50% acetonitrile-water(8.0 niL) at 0° C. After 1 h, the reaction mixture was loaded directlyonto a C₁₈-silica flash chromatography column and was eluted with 50%acetonitrile-water to provide synthetic (−)-saframycin A (1) as a yellowsolid (127 mg, 66%). Spectral data and chromatographic properties ofsynthetic 1 were identical to those of an authentic sample of naturalsaframycin A.

¹H NMR (500 MHz, CDCl₃), δ 6.67 (br dd, 1 H, J=8.1, 3.7 Hz, NHCO), 4.06(br d, 1 H, J=2.0 Hz, CHN(CH₃)CHCHC≡N), 4.02 (2 s, 3 H, each, 2×OCH₃),3.99 (d, 1 H, J=2.4 Hz, CHC≡N), 3.97 (m, 1 H, CHCH₂NH), 3.72 (ddd, 1 H,J=14.2, 8.8, 1.5 Hz, CH₂NH), 3.43 (br d, 1 H, J=7.6 Hz, CHCHC≡N), 3.26(dt, 1 H, J=14.2, 4.2 Hz, CH₂NH), 3.13 (dt, 1 H, J=11.4, 2.9 Hz,CHNCHC≡N), 2.88 (dd, 1 H, J=17.7, 2.6 Hz, CH₂CHCHNCH₃), 2.82 (dd, 1 H,J=21.1, 7.6 Hz, CH₂CHCHC≡N), 2.31 (s, 3 H, NCH₃), 2.25 (s, 3 H, COCH₃),2.24 (d, 1 H, J=21.0 Hz, CH₂CHCHC≡N), 1.98 (s, 3 H, CCH₃), 1.92 (s, 3 H,CCH₃), 1.28 (ddd, 1 H, J=17.7, 11.5, 2.7 Hz, CH₂CHNCHC≡N). ¹³C NMR (125MHz, CDCl₃), δ 196.7, 186.6, 185.3, 182.4, 180.8, 160.2, 155.9, 155.6,141.5, 141.2, 135.6, 135.5, 129.2, 128.3, 116.6, 61.1, 60.9, 58.2, 56.2,54.5, 54.2, 53.9, 41.6, 40.6, 25.0, 24.2, 21.5, 8.7 (2 C). FTIR (neatfilm), cm⁻¹ 3407 (w, NH), 2944 (w), 2249 (w, C≡N), 1720 (w), 1682 (m,C═O), 1652 (s, C═O), 1615 (m, C═C), 1447 (m). HRMS (ES) Calcd forC₂₉H₃₁N₄O₈ (M+H)⁺: 563.2142, Found: 563.2169. [α]_(D) ²³ (synthetic1)=−4.6°. [α]_(D) ³² (natural 1)=−4.0°.

B. In yet another embodiment of the present invention, the generalmethodology as described is exemplified by the one-step construction ofthe pentacyclic skeleton of saframycin A (9) from a trimeric α-aminoaldehyde precursor (8), as depicted in FIG. 3. Specifically, recognizingthat synthetic studies have shown that the potent antitumor alkaloidsaframycin A can be assembled from glycine, alanine, and two moleculesof tyrosine (Mikami et al. J. Biol. Chem. 1985, 260, 344; Arai et al.Antimicrob. Agents Chemother. 1985, 28, 5; Arai et al. In The Alkaloids;Brossi, A., Ed.; Academic Press: New York, 1983; Vol. 21, Chapter 3;Remers, W. A. In The Chemistry of Antitumor Antibiotics;Wiley-Interscience: New York, 1988; Vol. 2, Chapter 3.) As demonstratedabove (in part A), an efficient synthesis for the generation ofalkaloids via the directed condensation of α-amino aldehydes wasachieved. Over the course of the five steps described above, thecomponents were linked in a stepwise fashion in a sequence involving twoPictet-Spengler cyclization reactions and an intramolecular Streckerreaction, to form the pentacyclic saframycin A precursor 5 (same as (2)depicted in FIG. 1; see, Myers, A. G.; Kung, D. W. J. Am. Chem. Soc.1999, 121, 10828). In yet another embodiment, as demonstrated herein,alkaloid skeletons, specifically the saframycin skeleton as demonstratedbelow, can be assembled in one remarkable transformation from anN-linked oligomer of three cc-amino aldehyde components 2, 3 and 4,(FIG. 4) a reaction that suggests for the first time a viable pathwaylinking saframycin A (1) with an oligopeptide precursor and, therefore,a possible biosynthetic route.

The specific oligomer that was targeted initially was the trimeric aminonitrile, 6, in which 2, 3, and 4 are linked by sequential Streckerreactions (see FIG. 5). The amino nitrile groups serve to covalentlyjoin the three cc-amino aldehyde components and were proposed tofunction later as precursors to electrophilic imine or iminiumintermediates that would mediate the three cyclization reactions leadingto the saframycin skeleton (For selected examples of the use of aminonitrites as imine/iminium ion precursors in biomimetic systems, see (a)Overman, L. E.; Jacobsen, E. J. Tetrahedron Lett. 1982, 23, 2741 (b)Ksander et al. Helv. Chim. Acta 1987, 70, 1115; (c) Bonin et al. Org.Synth. 1992, 70, 54). Previously, it had been demonstrated that α-aminoaldehydes can be coupled using the Strecker reaction withoutepimerization of the α-stereocenter (Myers et al. J. Am. Chem. Soc.1999, 121, 8401). Because amino nitrile formation was anticipated toform two diastereomeric products in each case (of no consequence inlater C—C bond forming reactions) ¹³C labeled cyanide was utilized inthe synthesis to facilitate 13-C NMR analysis of the products. Also, thesequence was begun with a single diastereomer of the C-protected α-aminoaldehyde component 3, bearing a ¹³C-label on the cyano group.

The order of introduction of α-amino aldehyde components was 3 +2, then4, representing C- to N-terminus directionality in the synthesis. Mixing3 (1 equiv, 93% ee, ¹³C-labeled cyano group) and its N-protected α-aminoaldehyde counterpart 2 (1.05 equiv, 96% ee) in dichlorometane withsuspended sodium sulfate led to formation of the corresponding imine,cleanly and without α-epimerization, as previously demonstrated above.In this instance, however, the imine was captured by Strecker reactionwith hydrogen cyanide in methanol at 23° C. (1.6 equiv acetic acid, 1.5equiv K¹³CN, FIG. 5), whereas in the route described above, the iminewas cyclized by warming (35° C.) in the presence of lithium bromide. Theexpected α-amino nitriles 7 (1.1:1 mixture of diastereomers) wereobtained in 92% yield after isolation by flash column chromatograpy(FIG. 4). Sequential removal of the silyl ethers (triethylaminetrihydrofluoride, 2.5 equiv, CH₃CN, 23° C.) and the N-Fmoc group (30%piperidine-CH₂Cl₂, 23° C., 76%, two steps) of 7 afforded the fullydeprotected “dimer” for coupling with the third component, N-Fmocglycinal (4). Attempted Strecker coupling of these components wascomplicated by internal cyclization of the glycinaldimine intermediate.Recognizing that such a process provided an aminal product that wasfunctionally equivalent to the trimeric α-amino nitrile originallytargeted, the condensation reaction was optimized to form this product(compound 8, FIG. 5). Thus, addition of 4 (1.1 equiv) to a solution ofthe deprotected dimeric α-amino aldehyde (1 equiv) in dichloromethane at23° C. led to smooth condensation in the absence of hydrogen cyanide toafford a product formulated as the cyclic aminals 8. These products werenot stable to chromatography on silica gel, but ¹H- and 13C-NMR analysisshowed that they had been formed cleanly (˜90% combined yield). Onlydiastereomers were detected spectroscopically, and these were present inthe same ratio as the starting material 7, suggesting that the cyclicaminal has been formed with a single stereochemistry, tentativelyassigned as shown in FIG. 5.

Subsequently sequential treatment of 8 with the Lewis acids lithiumbromide (dimnethoxyethane, reflux) and then zinc chloride(trifluoroethanol-THF, 23° C.), and assisted by the fact that 8 wasfortuitously well separated chromatographically from all other reactionproducts, it was possible to isolate the desired pentacyclic saframycinA precursor 9 from the reaction mixture in pure form (4%). With furtherexperimentation, conditions were found to bring about the transformationof 8 to 9 in one step, and in higher yield (FIGS. 3 and 6); heating asolution of 8 in tetrahydrofuran at reflux in the presence of magnesiumbromide etherate (20 equiv) afforded 9 in 8.4 and 9.0% yield in twoseparate experiments. Importantly, N-acylation of 9 with theenantiomeric Mosher acid chlroides followed by HPLC analysis of theamide products established that 9 had been formed without racemization(9 was 99% ee). N-Methylation of 9 with formalin and sodiumtriacetoxyborohydride in acetonitrile afforded the pentacyclicsaframycin A precursor 5, (as depicted in FIG. 4) identical with anauthentic sample prepared by the earlier synthetic route (¹H NMR, IR,TLC, and HPLC analysis), except for the anticipated spectroscopicdifferences attributed to the ¹³C label. Intermediate 5 can betransformed into saframycin A in three steps (50% yield) (see, forexample, FIG. 2).

The one-step conversion of the N-linked oligomer 8 to the pentacyclicintermediate 9 involves an exceptional number of individual steps. Threecyclization reactions occur, and three of the five stereocenters ofsaframycin A are established in this step. In theory, each of the fivestereogenic centers of the precursor 5 (shown in FIG. 4) isepirmerizable under the reaction conditions. A single epimerizationevent may divert the course of reaction from 9. In that product which isformed, the α-amino aldehyde-derived centers are preserved. Many viablesequences can be envisioned to transform 8 into 9; the pathway shown inFIG. 6 is proposed as that which naturally occurs. In backgroundstudies, we have found that aminals have a greater propensity to formimine or iminium ion intermediates under mildly acidic conditions thansecondary amino nitrites which, in turn, are more labile than tertiaryamino nitriles (Myers et al. J. Am. Chem. Soc. 1999, 121, 8401). Forthis reason, without wishing to be bound by any particular theory, it isproposed that cleavage of the aminal occurs first, followed by trappingof the resultant imine by Pictet-Spengler cyclization, as depicted inFIG. 6. Subsequent ionization of the secondary amino nitrile is proposedto initiate a second Pictet-Spengler cyclization. Finally, ionization ofthe tertiary amino nitrile group leads to internal Strecker reaction toform the pentacyclic product 9. It is interesting to note that theordering of the two Pictet-Spengler reactions in this proposed sequenceis opposite to that of our earlier stepwise condensation route. BothPictet-Spengler cyclizations are believed to proceed with cisselectivity, as observed in the earlier stepwise route.

II. Synthesis of Analogues of saframycins:

A) General Procedures: All reactions were performed in oven-dried orflame-dried round-bottomed flasks. The flasks were fitted with rubbersepta or glass stoppers. Commercial reagents were used as received withthe following exceptions: THF was distilled from sodium benzophenoneketyl at 760 Torr, methanol was distilled from magnesium methoxide at760 Torr and dichloromethane was distilled from calcium hydride at 760Torr. The spectroscopic and analytical data for all of the analogs isdisclosed. All coupling constants are given in Hertz.

2) General Experimentals for Analogues:

It will be appreciated according to the novel methodology provided bythe present invention that useful compounds can be obtained as describedherein. In but one example, the methodology of the present inventionenables the rapid production of hydroquinones as shown below bearing aprotected amino functionality, which amino functionality can bedeprotected and reacted with suitable reagents under suitable reactionconditions, certain examples of which are described below, to generatederivatives, as described in more detail herein.

In certain embodiments of the present invention, acid chlorides bearingaryl, heteroaryl, aryloxy and alkyl functionalities are reacted with theamino compound presented above under suitable conditions withdiethylaniline in methylene chloride to generate desired compounds.

In certain other embodiments of the present invention, aldehydes bearingheteroaryl and aryl functionalities are reacted with the amino compoundpresented above under suitable conditions with NaBH(OAc)₃ inacetonitrile to generate desired compounds.

In still other embodiments of the present invention, carboxylic acidsbearing heteroaryl functionalities are reacted with the amino compoundpresented above under suitable conditions with EDC, HOBT, diethylanilinein tetrahydrofuran to generate desired compounds.

It will be appreciated by one of ordinary skill in the art that avariety of suitable reaction conditions can be utilized to functionalizethe amino moiety of the compounds depicted above, and thus thegeneration of the analogues as described herein is not intended to belimited to the specific examples described herein.

In addition to the syntheses of exemplary analogues from the aminofunctionality as described generally and depicted above, it will beappreciated that the method of the invention provides for the synthesisof pentacyclic structures bearing (at R₁) O, S and C-containingfunctionalities, thus enabling access to a variety of analogues.

It will be appreciated that the compounds as described herein can besynthesized using traditional solution phase methods (as describedgenerally above), or can be synthesized using solid-support techniques.

3. A Modular, Solid-Supported Synthesis of a Library of (−)-Saframycin AAnalogs

As described herein, compounds as described generally above and hereincan also be prepared using a modular solid-supported synthesis asdescribed herein. Additionally, as described herein, the modularsynthesis of the present invention permits extensive diversification ofthe core structure (e.g., R₁ moieties) as described herein. Furthermore,FIGS. 10-14B depict exemplary schemes and methods for the synthesis ofinventive compounds using solid-supported techniques.

Note on Resin Handling:

All solid-supported reactions were followed calorimetrically orchromatographically by directly sampling resin beads from the reactionsuspension via polypropylene syringe. The sampled beads were then washedon a polypropylene frit (Bio-Spin Disposable Chromatography Columns,P/N: 732-6008, Bio-Rad Laboratories, 2000 Alfred Nobel Drive, Hercules,Calif. 94547) and dried briefly in vacuo before being employed incalorimetric tests or exposed to cleavage cocktail (Kaiser, E., et. al.Anal. Biochem. 1976, 71, 261) (10 amine) and chloranil (Vojkovsky, T.Pept. Res., 1995, 8, 236) (2° amine)colorimetric assays were performedas reported (1999 Novabiochem Catalog & Peptide Synthesis Handbook,Calbiochem-Novabiochem Corporation, 10394 Pacific Center Court, SanDiego, Calif. 92121, pp. S43).

The progress of solid-supported reactions was followedchromatographically by liberating product from the solid support bymethanolysis of the siloxane linker: Washed resin samples (˜5 mg) weresuspended in a mixture of 100 IL dichloromethane, 20 μL methanol, and 10μL concentrated hydrochloric acid in a polypropylene Eppendorf tube andallowed to stand for 10 min at 23° C. with occasional manual agitation(˜every 3 min). The supernatant from this reaction mixture was analyzedby thin-layer chromatography, allowing semi-direct monitoring of thetransformation of the previously solid-immobilized compounds.Experimental Procedures:

Imidazole (7.33 g, 107.7 mmol, 1.1 equiv) was added in one portion to asolution of 5-hexen-1-ol (9.81 g, 97.91 mmol, 1.0 equiv) in 98.0 mLN,N-dimethylfornamide. The resulting clear solution was stirred for 10min at 0° C. and t-butyldimethylsilyl chloride (16.2 g, 107.7 mmol, 1.1equiv) was added. After stirring an additional 10 min at 0° C., theresulting reaction solution was allowed to warm to 23° C. and stirredfor 1.5 hrs. Excess t-butyldimethylsilyl chloride was then quenched bythe addition of 100 mL water, and the resulting aqueous solutionextracted with 2×200 mL diethyl ether. The combined organic extractswere then washed with 2×400 mL water and 1×500 mL brine, dried oversodium sulfate, and concentrated in vacuo to afford spectroscopicallypure siloxane product as a clear oil (22.28 g, 100%).

¹H NMR (500 MHz, CDCl₃), δ 5.85-5.77 (m, 1H, RCHCH₂), 5.00 (dq, 1H,J=17.5 Hz, RCHCH (Z)), 4.94 (dm, 1H, J=10.0 Hz, RCHCH (E)), 3.61 (t, 2H,J=6.5 Hz, CH₂OTBS), 2.06 (q, 2H, J=6.5 Hz, CH₂CHCH₂), 1.53 (m, 2H, J=6.0Hz, CH₂CH₂OTBS), 1.42 (m, 2H, J=6.0 Hz, CH₂CH₂CHCH₂), 0.90 (s, 9H,SiC(CH₃)₃), 0.02 (s, 6H, Si(CH₃)₂). ¹³C NMR (100 MHz, CDCl₃), δ 139.2,114.6, 63.3, 33.8, 32.5, 26.2, 25.4, 18.6, −5.1. FTIR (neat film), cm⁻¹3079 (w, CH), 2955 (s), 2930 (s), 2858 (s), 1472 (m), 1256 (s), 1103(s), 960 (m), 836 (s), 775 (s). R_(f) 0.70, 10% ethyl acetate-hexanes.HRMS (TOF-ES⁺) Calcd for C₁₂H₂₇OSi (M+H)⁺: 215.1831, Found.

3-chloroperoxybenzoic acid (442.0 mg, 77%, 1.97 mnmol, 1.2 equiv) wasadded in one portion to a solution of the siloxane substrate (352.2 mg,1.64 mmol, 1.0 equiv) in 8.0 mL dichloromethane at 0° C. After 5 min,the reaction solution was allowed to warm to 23° C. and stir for 13 hr.The reaction solution was then diluted with 80 mL pentane and washedsequentially with 1×80 mL sat. aq. sodium bicarbonate, 1×80 mL sat. aq.sodium bisulfite, 1×80 mL sat. aq. sodium bicarbonate, and 1×80 mLbrine. After drying over sodium sulfate, the organic layer wasconcentrated in vacuo and chromatographically purified (SiO₂7%ether-pentan→20% ether-pentane), providing the epoxide rac-3 as a clearoil (353.7 mg, 94%).

rac-3: ¹H NMR (400 MHz, CDCl₃), δ 3.62 (t, 2H, J=6.0 Hz, CH₂OTBS), 2.91(m, 1H, CH(O)CH₂), 2.75 (dd, 1H, J=4.0, 5.2 Hz, CH(O)CH₂ (Z)), 2.47 (dd,1H, J=2.8, 5.2 Hz, CH(O)CH₂ (E)), 1.61-1.47 (m, 6H, CH₂CH₂CH₂CH₂OTBS),0.98 (s, 9H, SiC(CH₃)₃), 0.05 (s, 6H, Si(CH₃)₂). 13C NMR (100 MHz,CDCl₃), δ 63.1, 52.5, 47.3, 32.8, 32.5, 26.2, 22.6, 18.6, −5.0. FTIR(neat film), cm⁻¹ 2951 (s), 2929 (s), 2856 (s), 1472 (s), 1255 (s), 1099(s), 836 (s), 775 (s). R_(f) 0.23, 5% ethyl acetate-hexanes. LRMS(TOF-ES⁺) Calcd for C₁₂H₂₇O₂Si (M+H)⁺: 231, Found: 231.

Glacial acetic acid (52.8 μL, 924.4 μmol, 10.2 equiv relative tocatalyst) was added to a red-orange solution of(S,S)-(+)-N,N′-Bis(3,5-di-tert-butylsalicylidene)-1,2-cyclohexanediaminocobalt(II)(52.9 mg, 87.62 μmol, 0.2 mol%) in 1.1 mL toluene at 23° C. in a flaskopen to the air. The resulting solution was stirred for 30 min at 23° C.and was then concentrated in vacuo to provide a brown solid. Epoxide(rac-3) (11.39 g, 43.40 mmol, 1.0 equiv) was added to this catalyst, andthe resulting brown-black solution stirred under argon at 0° C. for 10min before adding water (430.2 μL, 23.87 mmol, 0.55 equiv) to thereaction solution. The resulting mixture was stirred for 5 min at 0° C.before removing the cooling bath. After 28 hr at 23° C., theenantioenriched epoxide (S)-3 was distilled directly from the reactionmixture (95-99° C., 0.5 mm Hg), affording epoxide (S)-3 as a clear oil(4.62 g, 46%) and the product diol as a red-brown oil (5.39 g, 50%,contaminated with catalyst and water). Both the epoxide and the diolwere spectroscopically pure by ¹H NMR. Epoxide (S)-3 was determined tobe provided in >98% ee by Mosher ester analysis (¹⁹F NMR) of the 2°azidoalcohol provided by reaction of the epoxide with sodium azide.

(S)-3: ¹H NMR (400 MHz, CDCl₃), δ 3.62 (t, 2H, J=6.0 Hz, CH₂OTBS), 2.91(m, 1H, CH(O)CH₂), 2.75 (dd, 1H, J=4.0, 5.2 Hz, CH(O)CH₂ (cis-)), 2.47(dd, 1H, J=2.8, 5.2 Hz, CH(O)CH₂ (trans-)), 1.61-1.47 (m, 6H,CH₂CH₂CH₂CH₂OTBS), 0.98 (s, 9H, SiC(CH₃)₃), 0.05 (s, 6H, Si(CH₃)₂). ¹³CNMR (100 MHz, CDCl₃), δ 63.1, 52.5, 47.3, 32.8, 32.5, 26.2, 22.6, 18.6,−5.0. FTIR (neat film), cm⁻¹ 2951 (s), 2929 (s), 2856 (s), 1472 (s),1255 (s), 1099 (s), 836 (s), 775 (s). R_(f) 0.73, 50% ethylacetate-hexanes. LRMS (TOF-ES⁺) Calcd for C₁₂H₂₇O₂Si (M+H)⁺: 231, Found:231.

Diol: ¹H NMR (400 MHz, CDCl₃), δ 3.72 (m, 1H, CH₂CH(OH)CH₂), 3.66 (ddd,1H, J=3.2, 6.4, 10.8 Hz, CH₂OH), 3.63 (t, 2H, J=6.0 Hz, TBSOCH₂), 3.44(ddd, 1H, J=4.8, 7.6 Hz, 12.4 Hz, CH₂OH), 2.11 (m, 1H, CH₂CH(OH)), 1.86(m, 1H, CH₂CH(OH)), 1.51-1.58 (m, 2H, TBSOCH₂CH₂), 1.45-1.49 (m, 2H,CH₂CH₂CH(OH)), 0.89 (s, 9H, SiC(CH₃)), 0.05 (s, 6H, Si(CH₃)). ¹³C NMR(100 MHz, CDCl₃), δ 72.4, 67.0, 63.2, 33.1, 32.8, 26.2, 22.1, 18.6,−5.0. FTIR (neat film), cm⁻¹ 3372 (br, s, OH), 2935 (s, CH), 2857 (s,CH), 1472 (m), 1254 (m), 1102 (m), 836 (m), 775 (m). R_(f) 0.21, 50%ethyl acetate-hexanes. HRMS (TOF-ES⁺) Calcd for C₁₂H₂₉O₃Si (M+H)⁺:249.1886, Found: 249.1877.

A solution of expoxide (S)-3 (4.78 g, 20.75 mmol, 1.0 equiv) in 207 mLabs. EtOH was stirred at 0° C. for 10 min. Ethanolamine (62.6 mL, 1.04mol, 50.0 equiv), was then added gradually to the clear reactionsolution over 10 min, providing a yellow solution which was allowed towarm to 23° C. This solution was at 70° C. for 1 hr. Ethanol was removedfrom the reaction solution in vacuo and the resulting yellow oilpartitioned between 700 mL ethyl acetate and 700 mL water. The separatedorganic layer was then washed with 1×500 mL water and 1×500 mL brine,dried over potassium carbonate, and concentration in vacuo to providedanalytically pure aminodiol as a yellowish transparent oil (5.99 g,99%).

¹H NMR (400 MHz, CDCl₃), δ 3.68 (t, 2H, J=5.2 Hz, CH₂OH), 3.65 (m, 1H,CH(OH)), 3.62 (t, 2H, J=6.4 Hz, TBSOCH₂), 2.80 (q, 2H, J=5.6 Hz,CH₂CH₂OH), 2.74 (dd, 1H, J=3.2, 12.4 Hz, CH(OH)CH₂NH), 2.51 (dd, 1H,J=9.2, 12.0 Hz, CH(OH)CH₂NH), 1.50-1.58 (m, 2H, CH₂CH(OH)), 1.37-1.49(m, 4H, TBSOCH₂CH₂CH₂), 0.89 (s, 9H, SiC(CH₃)₃), 0.05 (s, 6H, Si(CH₃)₂).¹³C NMR (100 MHz, CDCl₃), δ 70.1, 63.3, 61.5, 55.2, 51.2, 35.0, 33.0,26.2, 22.2, 18.6, −5.0. FTIR (neat film), cm⁻¹ 3304 (br, s, OH), 2929(s), 2857 (s), 1471 (m), 1234 (m), 1098 (m). R_(f) 0.35, 50% ethylacetate-hexanes. HMS (TOF-ES⁺) Calcd for C₁₄H₃₄NO₃Si (M+H)⁺: 292.2308,Found: 292.2297.

Potassium bicarbonate (6.93 g, 69.17 mmol, 2.0 equiv) was added to asolution of substrate aminodiol (10.08 g 34.58 mmol, 1.0 equiv) in 300mL N,N-dimethylformamide. Benzyl bromide (4.20 mL, 34.58 mmol, 1.0equiv) was then added to the vigorously stirred resulting whitesuspension, the reaction vessal wrapped in foil, and the reaction vessalheated at 50° C. for 2.3 hr. After cooling, the reaction mixture waspartioned between 700 mL dichloromethane and 700 mL water and theorganic layer was separated. The aqueous layer was sequentiallyextracted with 1×500 mL, 1×300 mL, and 1×200 mL dichloromethane. All theorganic extracts were then combined, washed with 1×1.6 L water, anddried over sodium sulfate. Concentration of the dried extracts in vacuoprovided benzyl amine 4 as a viscous yellow oil which did not requirefurther purification (12.60 g, 92%).

4: ¹H NMR (400 MHz, CDCl₃), 1.3:1 mixture of rotamers, * indicates minorrotamer, δ 7.27-7.35 (m, 5H, C₆H₅), 3.868 (s, 1H*, CH₂*Ph), 3.83 (s, 1H,CH₂Ph), 3.64-3.71 (m, 2H, 2H*, CH₂OH, CH₂*OH), 3.59 (t, 2H, J=6.4 Hz,TBSOCH₂), 3.58 (s, 1H, CH₂Ph), 3.56 (s, 1H*, CH₂*Ph), 2.82 (dd, 1H*,CH₂*CH₂OH), 2.80 (dd, 1H, CH₂CH₂OH), 2.65 (t, 1H, CH₂CH₂OH), 2.61 (t,1H*, CH₂*CH₂OH), 2.56 (dd, 1H, J=3.2, 13.2 Hz, CH(OH)CH₂N), 2.47 (dd,1H, J=10.0, 12.8 Hz, CH(OH)CH₂N), 1.47-1.54 (m, 4H, TBSOCH₂CH₂CH₂CH₂),1.34-1.42 (m, 2H, TBSOCH₂CH₂CH₂), 0.89 (s, 9H, SiC(CH₃)₃), 0.04 (s, 6H,Si(CH₃)₂). ¹³C NMR (100 MHz, CDCl₃), δ 138.6, 129.0, 128.6, 127.4, 68.2,63.3, 61.0, 60.1, 59.8, 56.4, 34.8, 33.1, 26.3, 22.2, 18.7, −4.9. FTIR(neat film), cm⁻¹ 3362 (br, m, OH), 2924 (m), 2856 (m), 1460 (m), 1249(m), 1092 (m), 834 (m). R_(f) 0.34, 70% ethyl acetate-hexanes. HRMS(TOF-ES⁺) Calcd for C₂₁H₄₀NO₃Si (M+H)⁺: 382.2777, Found: 382.2760.

A solution of N-benzyl diol 4 (1.54 g, 4.02 mmol, 1.0 equiv) in 40.0 mLtetrahydrofuran was stirred at 0° C. for 10 min and was then added viacannula to sodium hydride (254.2 mg, 10.06 mmol, 2.5 equiv). Theresulting white suspension was stirred vigorously at 0° C. for 5 minbefore removing the cooling bath. After 1 hr at 23° C., the reactionsuspension was returned to a 0° C. bath for 10 min and N-tosylimidazole(894.5 mg, 4.02 mmol. 1.0 equiv) added in 3 portions over 12 min (gasevolution was observed subsequent to the addition of each portion).After stirring the resulting reaction solution for a further 10 min at0° C., the cooling bath was again removed. After 1 hr at 23° C., excesssodium hydride was CAREFULLY quenched by the slow addition of 30 mLsaturated aqueous ammonium chloride to the reaction suspension at 0° C.The reaction mixture was then partitioned between 340 mL anmouniumchloride (sat., aq.) and 370 mL diethyl ether and the organic layerseparated and washed further with 1×300 mL water and 1×200 mL brine. Theaqueous washes were combined and extracted with 2×200 mL diethyl ether.All of the organic extracts were then combined and dried over sodiumsulfate. Concentration of the extracts in vacuo provided a yellow oil,which was purified by flash column chromatography (SiO₂, 20% ethylacetate-hexanes), affording the N-benzyl morpholine product as a yellowoil (926.6 mg, 63%).

¹H NMR (400 MHz, CDCl₃), 1:1 mix of rotamers, * indicates rotamer, δ7.29-7.34 (m, 5H, C₆H₅), 3.83 (ddd, 1H, J=1.6, 2.8, 12.8 Hz, CHOCH₂),3.64 (dt, 1H, J=2.0, 11.2 Hz, CHOCH₂), 3.58 (t, 2H, J=6.0 Hz, TBSOCH₂),3.49 (s+m, 2H, CH₂Ph, CH(OR)CH), 3.48 (s, 1H*, CH₂*Ph), 2.72 (d, 1H,J=11.6 Hz, CH(OR)CH₂N), 2.65 (dd, 1H, J=1.6, 11.6 Hz, OCH₂CH₂N), 2.14(dt, 1H, J=3.2, 11.6 Hz, OCH₂CH₂N), 1.84 (t, 1H, J=11.2 Hz, CH(OR)CH₂N),1.41-1.54 (m, 4H, TBSOCH₂CH₂CH₂CH₂), 1.30-1.41 (m, 2H, TBSOCH₂CH₂CH₂),0.88 (s, 9H, SiC(CH₃)₃), 0.03 (s, 6H, Si(CH₃)₂). ¹³C NMR (100 MHz,CDCl₃), δ 129.2, 128.3, 127.2, 75.8, 67.0, 63.5, 63.3, 58.9, 53.4, 33.7,33.1, 26.2, 21.9, 18.7, −4.9. FTIR (neat film), cm⁻¹ 3372 (w), 2929 (s),2856 (s), 1454 (m), 1255 (m), 1101 (s), 835 (s). R_(f) 0.68, 70% ethylacetate-hexanes. LRMS (TOF-ES⁺) Calcd for C₂₁H₃₈NO₂Si (M+H)⁺: 364,Found: 364.

Glatial acetic acid (379.0 μL, 6.63 mmol, 2.6 equiv) was added to asolution of N-benzyl morpholine substrate (926.6 mg, 2.55 mmol, 1.0equiv) in 30.0 mL methanol. 10% Palladium on activated carbon (271.2 mg,254.8 mmol (Pd), 0.1 equiv), was then added to the clear solution. Theresulting black suspension was cycled under a H₂ atmosphere (1 atm) byalternately evacuating the reaction vessal and refilling with H2(g)(5×). After stirring 2 hr under 1 atm hydrogen, the reaction suspensionwas filtered through Celite 545 (CAUTION: do not allow the catalyst tobecome dry—ignition hazard), the catalyst washed with 3×100 mL methanol,and the filtrate partioned between 100 mL diethyl ether and 100 mLsodium bicarbonate (sat., aq.). The organic layer was then washed with afurther 1×80 mL brine and the combined aqueous washes extracted with1×100 mL diethyl ether. All organic extracts were then combined, driedover potassium carbonate, and concentrated in vacuo to providespectroscopically pure siloxymopholine 5 as an irridescent yellow-tingedoil (679.0 mg, 97%). Mosher amide analysis of siloxymorpoline 5 (¹H NMR)indicated that the product was provided in >95% ee.

5: ¹H NMR (400 MHz, CDCl₃), δ 3.87 (dt, 1H, J=2.0, 11.6 Hz, CHOCH₂),3.65 (ddd, 1H, J=7.2, 12.0, 14.0 Hz, CHOCH₂), 3.60 (t, 2H, J=6.4 Hz,TBSOCH₂), 3.47 (m, 1H, CH(OR)), 2.94 (dd, 1H, J=2.4, 12.4 Hz,CH(OR)CH₂), 2.88 (dd, 2H, J=2.8, 8.0 Hz, OCH₂CH₂N), 2.56 (dd, 1H,J=10.4, 12.4 Hz, CH(OR)CH₂N), 1.36-1.54 (m, 6H, TBSOCH₂CH₂CH₂CH₂), 0.88(s, 9H, SiC(CH₃)₃), 0.04 (s, 6H, Si(CH₃)₂). ¹³C NMR (100 MHz, CDCl₃), δ68.3, 63.2, 51.6, 46.2, 33.7, 33.0, 26.1, 21.7, 18.5, -5.1. FTIR (neatfilm), cm⁻¹ 3342 (w, NH), 2932 (s), 2856 (s), 1472 (m), 1461 (m), 1255(m), 1094 (s), 835 (s). R_(f) 0.43, 10% methanol-chloromethane,triethylamine-dipped plate. HRMS (TOF-ES⁺) Calcd for C₁₄H₃₂NO₂Si (M+H)⁺:274.2202, Found: 274.2191.

To a solution of siloxymorpholine 5 (606.6 mg, 2.22 mmol, 5.0 equiv) in4.5 mL 2,2,2-trifluoroethanol was stirred over a −20° C. bath for 10 minand was then added by cannula to substrate cyanohydrin (Prepared aspreviously reported: Myers, A. G.; Kung, D. W.; Zhong, B.; Movassaghi,M.; and Kwon, S. J. Am. Chem. Soc. 1999, 121, 8401-8402; Myers, A. G.;Kung, D. W. J. Am. Chem. Soc. 1999, 121, 10828-10829) (273.4 mg, 453.56μmol, 1.0 equiv) at −20° C. The resulting yellow solution was stirredfor 5 min at -20° C., then allowed to warm to 23° C. After 3 hr at 23°C., the reaction solution was concentrated in vacuo, then from 1×5.0 mLbenzene. Purification of the concentrate by flash column chromatography(SiO₂, 18% ethyl acetate-hexanes) afforded the syn-mopholinonitrile as aclear oil (149.6 mg, 38%) and the anti-mopholinonitrile as a clear oil(175.0 mg, 45%). Siloxymorpholine 5 was recovered from the column byelution with 10% methanol-chloromethane+2% (v/v) triethylamine,affording recovered siloxymorpholine 5 as a yellow oil (493.9 mg, 100%).

syn-morpholinonitrile (minor): ¹H NMR (400 MHz, CDCl₃), δ 7.76 (d, 2H,J=7.6 Hz, ArH), 7.54 (d, 2H, J=7.2 Hz, ArH), 7.40 (t, 2H, J=7.2 Hz,ArH), 7.30 (t, 2H, J=7.2 Hz, ArH), 6.54 (s, 1H, ArH), 5.29 (d, 1H, J=6.8Hz, NHFmoc), 4.38 (app. t, 1H, J=7.6 Hz, CO₂CH₂), 4.27 (app. t, 1H,J=10.8 Hz, CO₂CH₂), 4.20 (t, 1H, J=6.8 Hz, CO₂CH₂CH), 4.18 (br s, 1H,OCHCH₂), 3.86 (d, 1H, J=11.6 Hz, OCH₂CH₂), 3.74 (s, 3H, ArOCH₃), 3.71(s, 3H, ArOCH₃), 3.57 (t, 2H, TBSOCH₂), 3.50-3.40 (m, 2H, OCH₂CH₂,CHCN), 3.37 (m, 1H, CHNHFmoc), 3.10 (d, 1H, J=11.2 Hz, NCH₂), 2.94 (m,1H, NCH₂), 2.86 (d, 1H, J=11.2 Hz, NCH₂), 2.52 (d, 1H, 9.6 Hz, CH₂Ar),2.30 (t, 1H, J=10.4 Hz, CH₂Ar), 2.22 (s, 3H, ArCH₃), 1.52-1.26 (m, 6H,OCH₂CH₂CH₂CH₂), 0.99 (s, 9H, ArOSiC(CH₃)₃), 0.88 (s, 9H, ROSiC(CH₃)₃),0.16, 0.15 (2s, 6H, ArOSi(CH₃)₂), 0.03 (s, 6H, ROSi(CH₃)₂). ¹³C NMR (100MHz, CDCl₃), δ 151.5, 149.7, 145.4, 144.0, 141.5, 128.0, 127.3, 126.2,125.4, 125.2, 123.7, 121.0, 120.3, 115.0, 75.8, 67.0, 66.7, 63.2, 61.9,60.8, 60.1, 58.1, 50.9, 48.2, 47.3, 33.4, 33.0, 31.8, 26.2, 25.9, 21.7,18.6, 18.4, 10.2, −4.3, −5.0. FTIR (neat film), cm⁻¹ 3352 (w, NH), 2931(s), 2856 (s), 1727 (s, NCO₂),1482 (m), 1255 (s), 1238 (s), 1103 (m),1066 (s), 838 (s). R_(f) 0.53,30% ethyl acetate-hexanes. HRMS (TOF-ES⁺)Calcd for C₄₈H₇₂N₃O₇Si₂ (M+H)+: 858.4909, Found: 858.4890.

epi-syn-morpholinonitrile (trace): ¹H NMR (400 MHz, CDCl₃), δ 7.75 (d,2H, J=7.6 Hz, ArH), 7.57 (dd, 2H, J=2.8, 7.2 Hz, ArH), 7.39 (t,. 2H,J=7.6 Hz, ArH), 7.30 (t, 2H, J=7.2 Hz, ArH), 6.49 (s, 1H, ArH), 5.39 (d,1H, J=8.4 Hz, NHFmoc), 4.34-4.31 (m, 2H, CO₂CH₂), 4.20 (t, 1H, J=7.6 Hz,CO₂CH₂CH), 4.13 (m, 1H, OCHCH₂), 3.97 (d, 1H, J=10.8 Hz, ROCH₂CH₂), 3.72(s, 3H, ArOCH₃), 3.67 (s, 4H, ArOCH₃, CHCN), 3.61 (t, 3H, J=6.4 Hz,TBSOCH₂, ROCH₂CH₂), 3.53 (m, 1H, CHNHFmoc), 3.03 (dd, 1H, J=4.0, 14.0Hz, NCH₂), 2.87 (dd, 1H, J=7.6, 13.6 Hz, NCH₂), 2.81 (d, 1H, J=11.6 Hz,NCH₂), 2.57 (d, 1H, J=10.0 Hz, CH₂Ar), 2.52 (dd, 1H, J=2.8, 11.2 Hz,NCH₂), 2.34 (t, 1H, J=10.4 Hz, CH₂Ar), 2.21 (s, 3H, ArCH₃), 1.58-1.34(m, 6H, TBSOCH₂CH₂CH₂CH₂), 0.97 (s, 9H, ArOSiC(CH₃)₃), 0.89 (s, 9H,ROSiC(CH₃)₃), 0.12 (app. d, 6H, J=6.8 Hz, ArOSi(CH₃)₂), 0.05 (s, 6H,ROSi(CH₃)₂). FTIR (neat film), cm⁻¹ 3346 (w, NH), 2932 (s), 2856 (s),1721 (s, NCO₂), 1481 (m), 1451 (m), 1253 (s), 1104 (m), 1066 (s), 1010(m), 838 (s). R_(f) 0.49, 30% ethyl acetate-hexanes. HRMS (TOF-ES⁺)Calcd for C₄₈H₇₂N₃O₇Si₂ (M+H)⁺: 858.4909, Found: 858.4890.

anti-morpholinonitrile (major): ¹H NMR (400 MHz, CDCl₃), δ 7.76 (d, 2H,J=7.2 Hz, ArH), 7.60-7.52 (m, 2H, ArH), 7.40 (t, 2H, J=7.6 Hz, ArH),7.30 (dt, 2H, J=1.2, 7.6 Hz, ArH), 6.50 (s, 2H, ArH), 5.44 (d, 1H, J=8.0Hz, NHFmoc), 4.33 (d, 2H, J=7.6 Hz, CO₂CH₂), 4.21 (t, 1H, J=7.2 Hz,CO₂CH₂CH), 4.17 (m, 1H, OCHCH₂), 3.90 (d, 1H, J=10.0 Hz, OCH₂CH₂), 3.72(s, 3H, ArOCH₂), 3.67 (s, 3H, ArOCH₃), 3.63 (d, 1H, CHCN), 3.61 (t, 2H,J=6.4 Hz, TBSOCH₂), 3.55 (t, 1H, J=6.4 Hz, OCH₂CH₂), 3.51 (s, 1H,CHNFmoc), 3.02 (dd, 1H, J=4.0, 13.6 Hz, NCH₂), 2.89 (dd, 1H, J=7.6, 14.0Hz, NCH₂), 2.85 (d, 1H, J=10.0 Hz, NCH₂), 2.64 (dt, 1H, J=11.6 Hz,NCH₂), 2.52 (d, 1H, J=10.4 Hz, CH₂Ar), 2.25 (t, 1H, J=10.8 Hz, CH₂Ar),2.21 (s, 3H, ArCH₃), 1.56-1.36 (m, 6H, OCH₂CH₂CH₂CH₂), 0.98-0.96 (m, 9H,ArOSiC(CH₃)₃), 0.89-0.86 (m, 9H, ROSiC(CH₃)₃), 0.15-0.12 (m, 6H,ArOSi(CH₃)₂), 0.05-0.02 (m, 6H, ROSi(CH₃)₂). ¹³C NMR (100 MHz, CDCl₃), δ156.1, 151.4, 149.6, 145.3, 144.1, 141.4, 127.8, 127.2, 126.1, 125.3,124.3, 120.9, 120.8, 120.1, 75.8, 71.3, 67.4, 66.7, 63.2, 60.6, 60.0,53.1, 47.2, 33.6, 33.4, 33.0, 32.9, 31.7, 29.9, 26.1, 25.8(2), 21.8,18.3, 10.1, −4.4(2), −5.1. FTIR (neat film), cm⁻¹ 3341 (w, NH), 2929(s), 2857 (m), 1725 (s), 1481 (s), 1253 (s), 1237 (s), 1065 (s), 838(s). R_(f) 0.41, 30% ethyl acetate-hexanes. HRMS (TOF-ES+) Calcd forC₄₈H₇₂N₃O₇Si₂ (M+H)⁺: 858.4909, Found: 858.4873.

Concentrated hydrochloric acid (240.0 μL, 2.88 mmol, 1% v/v withmethanol, ˜6 equiv) was added to a solution of syn-aminonitrilesubstrate (406.2 mg, 473.3 plmol, 1.0 equiv) in 24.0 mL methanol thathad been stirred at 0° C. for 10 min. After 5 min, the reaction solutionwas partitioned between 240 mL diethyl ether and 240 mL (3:1)water:sodium bicarbonate (sat, aq). The organic layer was isolated andwashed with 1×50 mL brine. The combined aqueous layers were then furtherextracted with 1×250 mL diethyl ether and the combined organic extractsdried over sodium sulfate. Concentration in vacuo followed by columnchromatography (SiO₂, 60% ethyl acetate-hexanes) provided alcohol 7 as aclear oil (330.5 mg, 94%).

7: ¹H NMR (400 MHz, CDCl₃), δ 7.76 (d, 2H, J=8.0 Hz, ArH), 7.54 (d, 2H,J=7.6 Hz, ArH), 7.40 (t, 2H, J=8.0 Hz, ArH), 7.30 (t, 2H, J=7.6 Hz,ArH), 6.55 (s, 1H, ArH), 5.30 (app d, 1H, NHFmoc), 4.39 (m, 1H, OCH₂CH),4.27 (m, 1H, OCH₂CH), 4.20 (t, 1H, J=6.4 Hz, OCH₂CH), 3.87 (d, 1H,J=10.8 Hz, CH₂CHO), 3.74 (s, 3H, ArOCH₃), 3.72 (s, 3H, ArOCH₃), 3.66 (d,1H, J=6.8 Hz, CHCN), 3.61 (t, 2H, J=5.6 Hz, HOCH₂), 3.50 (d, 1H, J=11.2Hz, ROCH₂CH₂), 3.44 (d, 1H, J=9.2 Hz, ROCH₂CH₂), 3.37 (m, 1H, CHNHFmoc),3.09 (d, 1H, J=13.2 Hz, CH₂Ar), 2.93 (m, 1H, CH₂NCH₂), 2.87 (m, 1H,CH₂NCH₂), 2.52 (m, 1H, CH₂NCH₂), 2.42 (m, 1H, CH₂NCH₂), 2.32 (m, 1H,CH₂Ar), 2.22 (s, 3H, ArCH₃), 1.54-1.33 (m, 6H, HOCH₂CH₂CH₂CH₂), 0.99 (s,9H, C(CH₃)₃), 0.15, 0.16 (app d; 6H, Si(CH₃)₂). ¹³C NMR (100 MHz,CDCl₃), δ 175.2, 156.2, 151.5, 149.7, 144.0, 141.5, 128.0, 127.2, 126.2,125.2, 123.7, 120.9, 120.3, 115.9, 75.7, 67.1, 66.7, 62.9, 61.9, 60.8,60.1, 58.0, 50.9, 48.2, 47.3, 33.2, 32.8, 31.8, 25.9, 21.7, 184, 10.2,−4.3. FTIR (neat film), cm⁻¹ 3348 (br s, OH), 2934 (s), 2858 (s), 1721(s, NHCO₂), 1482 (m), 1238 (m), 1064 (m), 869 (m), 738 (m). R_(f) 0.50,70% ethyl acetate-hexanes. LRMS (TOF-ES⁺) Calcd for C₃₆H₄₃N₃NaO₇(M-TBS+Na)⁺: 651, Found: 651.

Concentrated hydrochloric acid (250.0 μL, 3.00 mmol, 1% v/v withmethanol, ˜3 equiv) was added to a solution of anti-aminonitrilesubstrate (916.2 mg, 1.07 mmol, 1.0 equiv) in 25.0 mL methanol that hadbeen stirred at 0° C. for 10 min. After 5 min, the reaction solution waspartitioned between 250 mL diethyl ether and 250 mL (3:1) water:sodiumbicarbonate (sat, aq). The organic layer was isolated and washed with1×150 mL brine. The combined aqueous layers were then further extractedwith 2×250 mL diethyl ether and the combined organic extracts dried oversodium sulfate. Concentration in vacuo followed by column chromatography(SiO₂, 60% ethyl acetate-hexanes) provided alcohol epi-7 as a clear oil(756.5 mg, 95%).

epi-7: ¹H NMR (400 MHz, CDCl₃), δ 7.76 (d, 2H, J=7.2 Hz, ArH), 7.58 (dd,2H, J=4.0, 7.2 Hz, ArH), 7.38 (t, 2H, J=7.6 Hz, ArH), 7.31 (dt, 2H,J=0.8, 7.2 Hz, ArH), 6.50 (s, 1H, ArH), 5.47 (d, 1H, J=8.8 Hz, NHFmoc),4.33 (d, 2H, J=7.2 Hz, CO₂CH₂), 4.21 (t, 1H, J=7.2 Hz, CO₂CH₂CH), 4.18(m, 1H, ROCH₂), 3.91 (d, 1H, J=9.6 Hz, OCHCH₂), 3.72 (s, 3H, ArOCH₃),3.67 (s, 3H, ArOCH₃), 3.66-3.61 (m, 4H, HOCH₂, ROCH₃, CHCN), 3.51 (m,1H, CHNHFmoc), 3.03 (dd, 1H, J=4.0, 14.0 Hz, NCH₂), 2.92-2.84 (m, 2H,NCH₂, CH₂Ar), 2.64 (dt, 1H, J=2.8, 10.8 Hz, NCH₂), 2.52 (d, 1H, J=10.8Hz, NCH₂), 2.27 (d, 1H, J=10.4 Hz, CH₂Ar), 2.21 (s, 3H, ArCH₃),1.61-1.36 (m, 6H, HOCH₂CH₂CH₂CH₂), 0.97 (s, 9H, SiC(CH₃)₃), 0.13 (app.d, 6H, J=4.4 Hz, Si(CH₃)₂). ¹³C NMR (100 MHz, CDCl₃), δ 156.2 151.5,149.7, 145.4, 144.2, 144.0, 141.5, 127.9, 127.3, 126.2, 125.5, 125.4,124.4, 121.0, 120.2, 115.6, 75.9, 67.5, 66.8, 62.9, 61.9, 60.7, 60.1,53.0, 52.7, 51.3, 47.3, 33.5, 32.8, 32.1, 25.9, 21.8, 18.4, 10.2, −4.3.FTIR (neat film), cm⁻¹ 3335 (m, NH/OH), 2933 (s), 2858 (m), 1714 (s,NCO₂), 1481 (s), 1451 (m), 1236 (s), 1117 (s), 1064 (m), 1012 (s), 867(s), 739 (m). R_(f) 0.46, 70% ethyl acetate-hexanes. HRMS (TOF-ES⁺)Calcd for C₄₂H₅₈N₃O₇Si (M+H)⁺: 744.4044, Found: 744.4060.

Polystyrene chlorodiisopropylsilane 6 was prepared by a modification ofthe procedure reported by Danishefsky (Randolph, J. T.; McClure, K. F.;Danishefsky, S. J. J. Am. Chem. Soc. 1995, 117, 5712-5719. Commercial4-bromopolystyrene (Novabiochem, 4-bromo polystyrene HL (P/N:01-64-0255), 50-100 mesh, Calbiochem-Novabiochem Corp., 10394 PacificCenter Court, San Diego, Calif. 92121) (3.65 g, 7.12 mmol, 1.95 mmol/g,1.0 equiv) was washed with 2×35 mL benzene under an argon atmosphere ina modified round-bottomed flask bearing an integral glass vacuum fritand was then resuspended in 35 mL benzene. n-Butyl lithium (2.50 M inhexanes, 7.72 mL, 19.29 mmol, 2.71 equiv) was added to this suspensionand the resulting mixture stirred at 60° C. for 3 hr. The cloudy whitereaction supernatant was then removed by vacuum filtration and the resinwashed with 1×35 mL benzene. The resin was then resuspended in 35 mLbenzene and dichlorodiisopropylsilane (5.14 mL, 28.47 mmol, 4.0 equiv)added. The resulting suspension was stirred for 3 hr at 23° C. and thereaction supernatant again removed via filtration. The resin was thensequentially washed with 2×36 mL (5:1) benzene:acetonitrile, 1×35 mLbenzene, 3×35 mL N,N-dimethylformamide, 2×35 mL tetrahydrofuran, and2×35 mL dichloromethane and dried in vacuo for 8 hr to providepolystyrene chlorodiisopropylsilane 6 as a yellowish-white, free-flowingresin (3.98 g).

A solution of alcohol 7 (54.1 mg, 72.7 μmol, 1.0 equiv) in 3.6 mLN,N-dimethylformamide was added via cannula to a mixture of polystyrenechlorodiisopropylsilane 6 (412.3 mg, 566.1 μmol, 7.8 equiv, 1.373mmol/g) and imidazole (44.6 mg, 654.4 μmol, 9.0 equiv) at 23° C. After 2hrs, additional imidazole (89.1 mg, 1.31 mmol, 18.0 equiv) and methanol(117.8 μL, 2.91 μmol, 40.0 equiv) were added to the resulting clearsuspension of yellowish resin and the resulting mixture stirred for 15hr. The reaction supernatant was then removed by crimped cannula, andthe product resin washed sequentially with 3×4 mL N,N-dimethylformamide,3×4 mL tetrahydrofuran, and 2×4 mL dichloromethane, removing the washsolution via crimped cannula Drying of the resin in vacuo for 5 hrprovided a free-flowing yellowish-white resin. The yield of thistransformation was determined by cleavage of the9-fluorenylmethoxycarbonyl (Fmoc) protecting group (20%piperidine-N,N-dimethylformamide, 2 min) from a measured aliquot ofresin followed by UV quantitation of the liberated dibenzofulvenechromophore (A₂₉₀) according to the established procedure (See, forexample, 1999 Novabiochem Catalog & Peptide Synthesis Handbook,Calbiochem-Novabiochem Corporation, 10394 Pacific Center Court, SanDiego, Calif. 92121, pp. S43). Averaging two replicate measurements gavea resin loading level of 0.1502 mmol/g (95%). (Note: trace Fmoc cleavageduring the loading and capping reactions suggests >95% loading.)Chromatographic separation of material cleaved from the product resin byincubating ˜5 mg samples of washed resin in a mixture of 100 μL CH₂Cl₂,20 μL CH₃OH, and 10 μL conc. hydrochloric acid for 10 min indicated thesolely the presence of substrate alcohol 7, confirming the cleanformation of product resin 8.

A solution of alcohol epi-7 (756.5 mg, 1.02 mmol, 1.0 equiv) in 40.0 mLN,N-dimethylformamide was added via cannula (along with 2×5.0 mLN,N-dimethylformamide washes) to a mixture of polystyrenechlorodiisopropylsilane 6 (3.98 g, 5.34 mmol, 5.24 equiv, 1.34 mmol/g)and imidazole (432.6 mg, 6.35 mmol, 6.25 equiv) at 23° C. After 5 hrs,additional imidazole (1.25 g, 18.3 mmol, 18.0 equiv) and methanol (1.65mL, 40.7 mmol, 40.0 equiv) were added to the resulting clear suspensionof yellowish resin and the resulting mixture stirred for 16 hr. Thereaction supernatant was then removed by crimped cannula, and theproduct resin washed sequentially with 3×40 mL N,N-dimethylformamide,3×40 mL tetrahydrofuran, and 2×40 mL dichloromethane, removing the washsolution via crimped cannula Drying of the resin in vacuo for 5 hrprovided a free-flowing yellowish-white resin. The yield of thistransformation was determined by cleavage of the9-fluorenylmethoxycarbonyl (Fmoc) protecting group (20%piperidine-N,N-dimethylformamide, 2 min) from a measured aliquot ofresin followed by UV quantitation of the liberated dibenzofulvenechromophore (A₂₉₀) according to the established procedure.⁷ Averagingtwo replicate measurements gave a resin loading level of 0.2345 mmol/g(100%). Chromatographic separation of material cleaved from the productresin by incubating ˜5 mg samples of washed resin in a mixture of 100 μLCH₂Cl₂, 20 μL CH₃OH, and 10 μL conc. hydrochloric acid for 10 minindicated the solely the presence of substrate alcohol epi-7, confirmingthe clean formation of product resin epi-8.

Acetic acid (253.8 μL, 4.44 mmol, 10.0 equiv) was added to a suspensionof aminonitrile resin 8 (2.12 g, 444.2 μmol, 1.0 equiv, 0.1575 mmol/g)in 20.0 mL tetrahydrofuran. Tetra-n-butylammmonium fluoride (1.0 M intetrahydrofuran, 2.22 mL, 2.22 mmol, 5.0 equiv) was then added to thereaction suspension and the resulting suspension stirred at 23° C. for1.5 hr. The reaction supernatant was then removed via crimped cannulaand the product resin washed sequentially with 3×20 mLN,N-dimethylformamide, 3×20 mL tetrahydrofuran, and 2×20 mLdichloromethane. The washed product resin was dried in vacuo overnight,affording a free-flowing yellow-orange resin. Photometric determinationof the resin 9-fluorenylmethoxycarbonyl (Fmoc) loading level (A₂₉₀, 20%piperidine-N,N-dimethylformamide) gave a resin loading level of 0.09504mmol/g, implying 45% Fmoc cleavage based on 100% yield for resin loadingstep. (TLC of the reaction supernatant confirms the formation ofdibenzofulvene, indicating that Fmoc deprotection does occur under thereaction conditions.) Substrate was liberated from the product resin forsolution-phase characterization by methanolysis of product resinaliquots (as described for 8 and epi-8 above), isolation of thesupernatant solution (cannulation) followed by resin washing, aqueousworkup of this substrate solution (sodium bicarbonate wash toneutralized hydrochloric acid), and chromatographic purification (90%ethyl acetate-hexanes).

¹H NMR (400 MHz, CDCl₃), δ 7.76 (d, 2H, J=7.6 Hz, ArH), 7.54 (d, 2H,J=7.2 Hz, ArH), 7.40 (t, 2H, J=7.6 Hz, ArH), 7.33 (t, 2H, J=6.0 Hz,ArH), 6.64 (s, 1H, ArH), 5.48 (s, 1H, ArOH), 5.32 (app d, 1H, J=6.4 Hz,NHFmoc), 4.32 (d, 2H, J=6.8 Hz, CO₂CH₂), 4.21 (t, 1H, J=6.8 Hz,CO₂CH₂CH), 4.16 (br s, 1H, CHOCH₂), 3.87 (d, 1H, J=10.8 Hz, CHOCH₂),3.77 (s, 3H, ArOCH₃), 3.73 (s, 3H, ArOCH₃),3.63 (m, 2H, HOCH₂),3.53-3.44 (m, 2H, CHCN, CHOCH₂), 3.39 (m, 11H, CHNHFmoc), 3.06 (d, 1H,J=13.6 Hz, OCH₂CH₂N), 2.96 (m, 1H, OCHCH₂N), 2.87 (d, 1H, J=10.0 Hz,OCH₂CH₂N), 2.55 (t, 1H, J=11.2 Hz, CH₂Ar), 2.43 (t, 1H, J=10.4 Hz,OCH₂CH₂N), 2.30 (t, 1H, J=8.4 Hz, CH₂Ar), 2.26 (s, 3H, ArCH₃), 1.60-1.30(m, 6H, HOCH₂CH₂CH₂CH₂). ¹³C NMR (100 MHz, CDCl₃), δ 156.2, 150.6,145.6, 145.4, 144.1, 143.9, 141.5, 128.0, 127.3, 125.3, 125.1, 120.3,116.0, 115.3, 75.7, 67.1, 66.6, 63.0, 61.8, 61.0, 60.9, 57.8, 51.1,48.7, 47.3, 33.2, 32.7, 31.6, 21.7, 10.3. FTIR (neat film), cm⁻3347 (m,NH), 2939 (s), 2864 (m), 1712 (s, NCO₂), 1483 (s), 1450 (s), 1233 (s),1111 (s), 1051 (s), 1008 (m), 738 (s). R_(f) 0.14, 70% ethylacetate-hexanes. HRMS (TOF-ES+) Calcd for C₃₆H₄₄N₃O₇ (M+H)⁺: 630.3179,Found: 630.3192.

Acetic acid (9.0 μL, 156.8 μmol, 2.2 equiv) and tetra-n-butylammoniumfluoride (1.0 M in tetrahydrofuran, 78.4 μL, 78.4 μmol, 1.1 equiv) weresequentially added to a suspension of siloxane resin epi-8 (328.0 mg,71.27 μmol, 1.0 equiv) in 3.5 mL tetrahydrofuran that had been stirringat 0° C. for 10 min. After 4.5 hr, the reaction supernatant was removedvia crimped cannula and the product resin washed with 2×4 mLN,N-dimethylformamide, 2×4 mL tetrahydrofuran, and 2×4 mLdichloromethane. The product resin was then dried in vacuo for 5 hr toprovide a free-flowing, yellow resin. (Note: TLC of the reactionsupernatant reveals the formation of dibenzofulvene, indicating thatFmoc deprotection occurs under the reaction conditions (seecorresponding transformation of 8 above).)

N-Fmoc-morpholinonitrile substrate resin (2.06 g, 443.5 μmol, 1.0 equiv,0.2152 mmol/g) was suspended in 20.0 mL 20% (v/v)piperidine-N,N-dimethylformamide. The resulting suspension was stirredat 23° C. for 1.5 hr and the reaction supernatant then removed viacrimped cannula The product resin was then sequentially washed with 3×20mL N,N-dimethylformamide, 3×20 mL tetrahydrofuran, and 2×20 mLdichloromethane and dried in vacuo to provide product resin 9 as afree-flowing tan solid. Methanolysis of aliquots of washed product resinindicated the sole presence of the expected siloxane cleavage product.

9: ¹H NMR (400 MHz, CDCl₃), δ 6.68 (s, 1H, ArH), 3.93 (ddd, 1H,J=1.6,3.6, 11.2 Hz, CHOCH₂), 3.78 (s, 3H, ArOCH₃), 3.70 (s, 3H, ArOCH₃),3.64 (t, 2H, J=6.4 Hz, HOCH₂), 3.59 (dt, 1H, J=2.8, 11.6 Hz, CHOCH₂),3.51 (m, 1H, CHNH₂), 3.38 (app dd, 1H, J=2.8, 8.4 Hz, CHOCH₂), 3.24 (d,1H, J=10.0 Hz, CHCN), 3.07 (dd, 1H, J=3.2, 13.2 Hz, OCHCH₂N), 2.64 (d,1H, J=11.2 Hz, OCH₂CH₂N), 2.56 (t, 1H, J=12.4 Hz, CH₂Ar), 2.53 (dd, 1H,J=6.4, 11.6 Hz, OCH₂CH₂N), 2.48 (dt, 1H, J=3.6, 11.6 Hz, OCH₂CH₂N), 2.41(t, 1H, J=10.8 Hz, CH₂Ar), 2.24 (s, 1H, ArCH₃), 1.62-1.35 (m, 6H,HOCH₂CH₂CH₂CH₂). ¹³C NMR (100 MHz, CDCl₃), δ 150.7, 145.5, 145.2, 126.3,125.2, 115.8, 114.8, 75.9, 66.5, 64.5, 62.9, 60.9 (2), 58.3, 50.3, 47.2,34.9, 33.2, 32.8, 21.7, 10.2. FTIR (neat film), cm⁻¹ 3356 (br, m, NH,OH), 2940 (s, CH), 2862 (m), 1483 (m), 1458 (m), 1420 (m), 1113 (s),1051 (m), 1010 (m), 911 (m), 732 (s). R_(f) 0.28, 10%methanol-dichloromethane, triethylamine-dipped plate. HRMS (TOF-ES⁺)Calcd for C₂₁H₃₄N₃O₅ (M+H)⁺: 408.2498, Found: 408.2516.

N-Fmoc-morpholinonitrile substrate resin (315.0 mg, 70.19 μmol, 1.0equiv, 0.2228 mmol/g) was suspended in 3.95 mL 20% (v/v)piperidine-N,N-dimethylformamide. The resulting suspension was stirredat 23° C. for 5 hr and the reaction supernatant then removed via crimpedcannula. The product resin was then sequentially washed with 3×4 mLN,N-dimethylformamide, 3×4 mL tetrahydrofuran, and 2×4 mLdichloromethane and dried in vacuo to provide product resin epi-9 as afree-flowing cream-colored resin. Solid-immobilized substrate wasliberated for characterization by a suspending a sample of the productresin (179.0 mg, 41.96 μmol, 1.0 equiv) in a mixture of dichloromethane(5.0 mL), methanol (170.0 μL, 4.2 mmol, 100.0 equiv), and concentratedhydrochloric acid (17.1 μL, 205.6 μmol, 4.9 equiv) at 23° C. over 22 hr.The reaction supernatant from this mixture was collected via crimpedcannula along with 6×4 mL dichloromethane washes of the treated productresin sample and the combined filtrates partitioned between 30 mL sat.aqueous sodium bicarbonate and 30 mL dichlormethane. The organic layerwas isolated and the aqueous layer extracted with a further 6×30 mLdiethyl ether. The organic extracts were then combined, dried oversodium sulfate, and concentrated in vacuo. Chromatographic purificationof the resulting orange oil (SiO₂, 3% methanol-dichlormethane on acolumn packed in eluent+5% triethylamine) provided the expected cleavedamine substrate as a yellow-tinged clear oil (5.2 mg, 28%).

epi-9: ¹H NMR (400 MHz, CDCl₃), δ 6.61 (s, 1H, ArH), 3.91 (dm, 1H,J=11.6 Hz, ROCH₂), 3.79 (s, 3H, ArOCH₃), 3.72-3.67 (m, 3H, ROCH₂,OCHCH₂, CHCN), 3.66 (s, 3H, ArOCH₃), 3.52-3.44 (m, 2H, HOCH₂), 3.32 (dt,1H, J=3.2, 8.4 Hz, CHNH₂), 3.16 (dd, 1H, J=3.2, 13.2 Hz, NCH₂), 2.81 (d,1H, J=10.8 Hz, CH₂Ar), 2.65 (dt, 1H, J=3.6, 11.2, NCH₂), 2.54 (d, 1H,J=10.4 Hz, NCH₂), 2.37 (dd, 1H, J=8.4, 13.6 Hz, NCH₂), 2.24 (s, 3H,ArCH₃), 2.22-2.15 (m, 1H, CH₂Ar), 1.64-1.40 (m, 6H, HOCH₂CH₂CH₂CH₂), ¹³CNMR (100 MHz, CDCl₃), δ 150.7, 145.5, 145.0, 127.2, 125.0, 116.4, 114.6,75.7, 66.7, 65.9, 63.0, 60.9, 53.3, 52.8, 51.4, 35.2, 33.4, 32.8, 21.8,17.4, 10.2. FTIR (neat film), cm⁻¹ 3358 (m, NH₂/OH), 2936 (s), 2864 (m),1483 (m), 1452 (m), 1417 (m), 1232 (w), 1112 (s), 1050 (m), 1009 (m).R_(f) 0.34, 10% methanol-dichloromethane, triethylamine-dipped plate.HRMS (TOF-ES⁺) Calcd for C₂₁H₃₄N₃O₅ (M+H)⁺: 408.2498, Found: 408.2480.

A solution of aldehyde Prepared as previously reported: (Myers, A. G.;Kung, D. W.; Zhong, B.; Movassaghi, M.; and Kwon, S. J. Am. Chem. Soc.1999, 121, 8401-8402) (726.6 mg, 1.26 mmol, 2.84 equiv) in 15.0 mLN,N-dimethylformamide was added to amine resin 9 (1.96 g, 443.5 μmol,1.0 equiv, 0.2261 mmol/g) via cannula using 2×5 mL N,N-dimethylformamidewashes to quantitate the transfer. The resulting clear suspension oforangish resin was stirred at 23° C. in the dark for 5 hr before thereaction supernatant was removed via crimped cannula and the productresin washed sequentially with 2×20 mL N,N-dimethylformamide and 2×20 mL1,2-dimethoxyethane. (Note: the reaction supernatant andN,N-dimethylforrnamide washes were collected to allow recovery of excessaldehyde (see below).) The resulting yellow resin (10) was employeddirectly in the synthesis of tetrahydroisoquinoline resin 11.

A solution of anhydrous lithium bromide (3.41 g, 39.30 mmol, 88 equiv)in 30.0 mL 1,2-dimethoxyethane (prepared with gentle heating) was addedto imine resin 10 (2.21 mg, 443.5 μmol, 1.0 equiv, 0.2008 mmol/g) via awarm syringe and needle. The resulting clear suspension of yellow resinwas placed on a 35° C. bath and stirred for 22 hr. The supernatant wasthen removed via crimped cannula and the product resin washedsequentially with 4×20 mL N,N-dimethylformamide, 3×20 mLtetrahydrofuran, and 2×20 mL dichloromethane and dried in vacuo toprovide free-flowing orangish-white resin which gave a negative Kaisertest and positive chloranil test. Photometric determination of theproduct resin 9-fluorenylmethoxycarbonyl (Fmoc) loading level (A₂₉₀, 20%piperidine-N,N-dimethylformamide, duplicate measurements) established aresin loading level of 0.1698 mmol/g, consistent with an 81% yield ofdiastereomeric tetrahydroisoquinolines (average 95% yield/step, 4steps). Liberation of substrate for characterization and establishmentof diastereomeric ratio for the transformation was effected byincubating a suspension of product resin 11 (86.8 mg, 13.2 μmol (basedon Fmoc loading level), 1.0 equiv) in 2.0 mL dichloromethane withmethanol (53.5 μL, 1.32 mmol, 100.0 equiv) and concentrated hydrochloricacid (5.4 μL, 64.8 μmol, 4.9 equiv) at 23° C. After 1 hr, the reactionsupernatant was collected via crimped cannula, along with 2×2 mLdichloromethane washes of the treated resin sample. This filtrates werepartitioned between 30 mL dichloromethane and 30 mL sat. aq. sodiumbicarbonate, the organic layer isolated, and the aqueous layer extractedwith a further 2×30 mL diethyl ether. The combined organic extracts werethen dried over sodium sulfate, concentrated in vacuo, andchromatographically purified (SiO₂, 70% ethyl acetate-hexanes→80% ethylacetate-hexanes→100% ethyl acetate-hexanes), affording cis-11 (3.4 mg,27%) and a mixture of trans-11 and epi-11 (epimeric at the aminonitrilemethine and both centers in the tetrahydroisoquinoline ring) (0.8 mg,6.3%, 1.8:1 trans- to epi-) as white foams (˜6.7:1 diastereomeric ratioof cis- and trans-products).

cis-11: ¹H NMR (500 MHz, CDCl₃), * denotes non-fused aromatic ringprotons, δ 7.74 (d, 2H, J=8.0 Hz, ArH), 7.49 (d, 1H, J=7.0 Hz, ArH),7.43 (d, 1H, J=7.5 Hz, ArH), 7.40-7.36 (m, 2H, ArH), 7.29-7.25 (m ,2H,ArH), 6.36 (s, 1H, ArH), 6.18 (s, 1H, ArOH), 5.68 (d, 1H), J=6.5 Hz,NHFmoc), 4.82 (s, 1H, NHCH), 4.57 (br s, 1H, NCHAr), 4.43 (dd, 1H,J=7.0, 10.5 Hz, CO₂CH₂), 4.16 (t, 1H, J=7.5 Hz, CO₂CH₂CH), 4.09 (app. t,1H, J=10.0 Hz, CO₂CH₂), 3.93 (d, 1H, J=10.5 Hz, ROCH₂CH₂), 3.79 (s, 3H,ArOCH₃*), 3.69 (s, 3H, ArOCH₃*), 3.65 (s, 6H, 2×ArOCH₃), 3.61-3.58 (m,1H, ROCH₂CH₂), 3.57 (app. s, 2H, HOCH₂), 3.47 (d, 1H, J=10.5 Hz, CHCN),3.37 (br s, 1H, OCHCH₂), 3.20 (dd, 1H, J=1.5, 14.5 Hz, CH₂Ar), 3.11 (t,1H, J=10.5 Hz, CH(CN)CH), 2.87 (t, 1H, J=13.5 Hz, CHNHFmoc), 2.65 (t,2H, J=11.0 Hz, 2□NCH₂), 2.45 (t, 2H, J=11.0 Hz, 2×NCH₂), 2.33 (dd, 1H,J=11.0, 14.0 Hz, CH₂Ar), 2.25 (s, 3H, ArCH₃*), 2.19 (s, 3H, ArCH₃), 2.12(t, 2H, J=13.0 Hz, CH₂Ar*), 1.64-1.40 (m, 6H, HOCH₂CH₂CH₂CH₂), 0.96 (s,9H, SiC(CH₃)₃), 0.10 (2×s, 6H, Si(CH₃)_(s)). ¹³C NMR (100 MHz, CDCl₃), δ156.7, 149.3, 148.8, 145.9, 145.3, 144.5, 144.1, 144.0, 142.8, 141.5,140.1, 127.9, 127.3, 125.4, 124.8, 122.6, 120.3, 120.2, 119.8, 115.0,75.8, 67.1, 66.5, 64.2, 63.0, 60.9, 60.1, 58.3, 56.8, 55.2, 52.2, 50.5,47.4, 47.2, 33.3, 32.9, 29.9, 29.2, 28.6, 25.9, 21.8, 18.4, 10.1, 9.9,−4.4. FTIR (neat film), cm⁻¹ 3344 (m, NH), 2934 (s), 2857 (m), 1714 (s,NCO₂), 1463 (s), 1452 (s), 1238 (s), 1113 (s), 1060 (s), 869 (m), 783(m). R_(f) 0.48, 80% ethyl acetate-hexanes. HRMS (TOF-ES⁺) Calcd forC₅₄H₇₃N₄O₁₀Si (M+H)⁺: 965.5096, Found: 965.5127.

The N,N-dimethylformamide solution of substrate aldehyde isolated byfiltration of resin 10 (see above) was partitioned between 250 mLdiethyl ether and 250 mL water and the ether layer isolated. The aqueouslayer was extracted further with 2×250 mL (1:1) hexanes-diethyl etherand all organic extracts combined, dried over sodium sulfate, andconcentrated in vacuo (some N,N-dimethylformamide remained). Theresulting oil (471.2 mg aldehyde (theoretical), 818.5 μmol, 1.0 equiv)was dissolved in 27.0 mL methanol and stirred at 0° C. for 10 min beforeadding sodium borohydride in one portion (15.5 mg, 409.2 μmol, 0.5equiv). After 25 min at 0° C., the reaction solution was diluted with 15mL diethyl ether and excess sodium borohydride carefully quenched by theSLOW addition of 15 mL saturated aqueous ammonium chloride. Theresulting solution was partitioned between 150 mL diethyl ether and 150mL saturated aqueous ammonium chloride and the organic layer separatedand washed with 1×100 mL sat. aq. ammonium chloride and 1×100 mL brine.All combined aqueous washes were then extracted 1×250 mL diethyl etherand all organic extracts were then combined, dried over sodium sulfate,and concentrated in vacuo. Chromatographic purification of the resultingproduct (SiO₂, 70% ethyl acetate-hexanes) provided the previouslyreported alcohol product (Myers, A. G.; Kung, D. W.; Zhong, B.;Movassaghi, M.; and Kwon, S. J. Am. Chem. Soc. 1999, 121, 8401-8402) asa clear oil (458.7 mg, 97%).

Formalin (660.5 μL, 8.87 mmol, 20.0 equiv) was added to a suspension oftetrahydroisoquinoline resin 11 (2.21 mg, 443.5 mmol, 1.0 equiv, 0.1762mmol/g) and sodium triacetoxyborohydride (1.41 g, 6.65 mmol, 15.0 equiv)in 40.0 mL N,N-dimethylformamide. The resulting clear suspension oforangish resin was stirred for 3.5 hr before removing the reactionsupernatant via crimped cannula and washing the resin sequentially with5×30 mL N,N-dimethylformamide, 5×30 mL tetrahydrofuran, and 2×30 mLdichloromethane. Drying the product resin in vacuo provided afree-flowing, light orange resin than gave a negative chloranil testPhotometric determination of the product resin9-fluorenylmethoxycarbonyl (Fmoc) loading level (A₂₉₀, 20%piperidine-N,N-dimethylformamide, duplicate measurements) established aresin loading level of 0.1571 mmol/g indicating a 95% yield for thistransformation given chromatographic and colorimetric evidence for thecompletion of this reaction. Substrate cleavage for characterization waseffected by incubating product resin (95.5 mg, 16.8 μmol, 1.0 equiv) in2.7 mL dichloromethane with methanol (68.9 μL, 1.70 mmol, 100.0 equiv)and concentrated hydrochloric acid (7.0 μL, 83.3 μmol, 4.9 equiv) for 18hr at 23° C. The reaction supernatant was then removed by crimpedcannula and collected along with 5×2.5 mL dichloromethane washes of thetreated resin. These filtrates were partitioned between 30 mLdichloromethane and 30 mL sat. aq. sodium bicarbonate and the organiclayer was isolated. After further extraction of the aqueous layer (2×30mL diethyl ether), all organic extracts were combined, dried over sodiumsulfate, and concentrated in vacuo. Chromatographic purification of theresulting oil (prep TLC, SiO₂, 5% methanol-dichloromethane) provided thecis-tetrahydroisoquinoline product (10.2 mg, 62%) andtrans-tetrahydroisoquinoline product (1.5 mg, 9%) as white solids (6.8:1diastereomeric ratio).

cis-amine: ¹H NMR (500 MHz, CDCl₃), * denotes non-fused aromatic ringprotons, δ 7.72 (dd, 2H, J=4.0, 7.5 Hz, ArH), 7.41 (t, 2H, J=7.5 Hz,ArH), 7.36 (t, 2H, J=7.5 Hz, ArH), 7.26 (m, 2H, ArH), 6.52 (s, 1H, ArH),6.17 (br s, 1H, ArOH), 5.50 (d, 1H, J=8.5 Hz, NHFmoc), 4.17-4.14 (m, 1H,CO₂CH₂), 4.00 (t, 1H, J=7.5 Hz, CO₂CH₂CH), 3.98-3.94 (m, 1H, CO₂CH₂),3.93 (br s, 1H, NCHAr), 3.75 (d, 1H, J=8.0 Hz, ROCH₂CH₂), 3.68 (2×s,(6H, 2×ArOCH₃*), 3.66 (s, 3H, ArOCH₃), 3.65 (s, 3H, ArOCH₃), 3.60 (s,2H, HOCH₂), 3.54 (m, 1H, ROCH₂CH₂), 3.48 (dt, 1H, J=2.0, 6.5 Hz,OCHCH₂), 3.43 (s, 1H, CHCN), 3.31 (d, 1H, J=11.0 Hz, CH₂Ar), 2.96-2.93(m, 1H, CHNCH₃), 2.93-2.88 (m, 1H, CHNHFmoc), 2.75-2.67 (m, 3H, 2□NCH₂,CH₂Ar), 2.59 (s, 3H, NCH₃), 2.52 (app. dt, 2H, J=2.5, 11.0 Hz, NCH₂,CH₂Ar*), 2.26-2.19 (m, 2H, NCH₂, CH₂Ar*), 2.19 (s, 3H, ArCH₃*), 2.17 (s,3H, ArCH₃), 1.61-1.49 (m, 4H, HOCH₂CH₂CH₂CH₂), 1.45-1.35 (m, 2H,HOCH₂CH₂CH₂), 0.95 (s, 9H, SiC(CH₃)₃), 0.11 (app. d, 6H, J=5.5 Hz,Si(CH₃)₂). ¹³C NMR (100 MHz, CDCl₃), δ 156.4 151.6, 148.7, 148.5, 145.1,144.3 (2), 144.1, 142.6, 141.3 (2), 127.8, 127.2 (2), 126.9, 125.4,125.3, 125.2, 123.7, 122.9, 120.6, 120.1, 120.0, 116.3, 75.8, 66.9,66.6, 64.7, 63.9, 63.7, 63.0, 61.4, 61.2, 60.8, 60.0, 58.4, 57.9, 50.5,47.4, 33.3, 32.8, 31.2, 29.9, 25.9, 24.7, 21.8, 18.4, 10.1, 9.8, −4.4.FTIR (neat film), cm⁻¹ 3390 (m, NH), 2932 (s), 2857 (m), 1708 (s, NCO₂),1480 (m), 1450 (m), 1416 (m), 1237 (m), 1115 (m), 1061 (s), 1009 (m),870 (m), 741 (m). R_(f) 0.42, 5% methanol-dichloromethane. HRMS(TOF-ES⁺) Calcd for C₅₅H₇₅N₄O₁₀Si (M+H)⁺: 979.5252, Found: 979.5210.

Tetra-n-butylammonium fluoride (1.0 M in tetrahydrofuran, 2.03 mL, 2.03mmol, 5.0 equiv) was added to a suspension of N-methylated amine resin(2.42 g, 405.4 μmol, 1.0 equiv, 0.1678 mmol/g) and glatial acetic acid(231.7 μL, 4.05 mmol, 10.0 equiv) in 20.0 mL tetrahydrofuran at 23° C.The resulting suspension was stirred for 2.5 hr before the reactionsupernatant was removed via crimped cannula. The product resin waswashed sequentially with 2×20 mL N,N-dimethylformamide, 3×20 mLtetrahydrofuran, and 2×20 mL dichloromethane and dried in vacuo toprovide a free-flowing yellow-orange resin. Cleaved substrate wasrecovered from the isolated reaction supernatant andN,N-dimethylformamide washes to determine the extent of siloxanecleavage through this procedure. The washes and supernatant wereconcentrated in vacuo to ˜3 mL then partitioned between 30 mL diethylether and 30 mL (3:1) water-sodium bicarbonate (sat., aq.). The organiclayer was separated and the aqueous layer extracted with 2×30 mL diethylether. All organic extracts were combined, dried over sodium sulfate,concentrated in vacuo, and chromatographically purified (prep TLC, 7%methanol-dichloromethane) to provide the expected siloxane cleavageproducts, the cis-tetrahydroisoquinoline (4.5 mg, 1.3%), and presumedtrans-tetrahydroisoquinoline (0.5 mg, ˜0.14%), as white solids.Photometric determination of the untreated product resin9-fluorenylmethoxycarbonyl (Fmoc) loading level (A₂₉₀, 20%piperidine-N,N-dimethylformamide, duplicate measurements) established aresin loading level of 0.07732 mmol/g, suggesting that ˜58% of theresin-bound product had lost its Fmoc protecting group in thistransformation (assuming a 98% yield for the phenolic siloxanedeprotection (based on substrate resin-cleavage yields)). Thisdeprotection of resin-bound product was confirmed by the isolation ofdibenzofulvene (13.2 mg, 33%) from the reaction supematant along withthe diastereomeric tetrahydroisoquinolines described above.

cis-tetrahydroisoquinoline: ¹H NMR (500 MHz, CDCl₃), * denotes non-fusedaromatic ring protons, δ 7.72 (d, 2H, J=7.2 Hz, ArH), 7.43 (d, 2H, J=8.0Hz, ArH), 7.36 (q, 2H, J=7.2 Hz, ArH), 7.26 (app. q, 2H, ArH), 6.64 (s,1H, ArH), 6.21 (br s, 1H, ArOH), 5.63 (d, 1H, J=8.4 Hz, NHFmoc), 5.34(br s, 1H, ArOH*), 4.24 (dd, 1H, J=6.8, 10.4 Hz, CO₂CH₂), 4.16 (d, 1H,J=6.0 Hz, CO₂CH₂), 4.03 (t, 1H, J=7.2 Hz, CO₂CH₂CH), 3.98-3.89 (m, 2H,NCHAr, ROCH₂CH₂), 3.74 (d, 1H, J=4.8 Hz, ROCH₂CH₂), 3.70 (s, 3H,ArOCH₃*), 3.69 (s, 3H, ArOCH₃*), 3.68 (s, 3H, ArOCH₃), 3.66 (s, 3H,ArOCH₃), 3.62 (m, 1H, OCHCH₂), 3.54 (br s, 2H, HOCH₂), 3.48 (m, 1H,CHCN), 3.31 (d, 1H, J=11.6 Hz, CH₂Ar), 3.03 (dd, 1H, J=10.8, 13.6 Hz,CHNCH₂), 2.93 (d, 1H, J=12.0 Hz, CHNHFmoc), 2.83 (dd, 1H, J=11.2 Hz,CH₂Ar), 2.75-2.66 (m, 2H, 2×NCH₂), 2.59 (s, 3H, NCH₃), 2.53 (dt, 2H,J=2.8, 12.4 Hz, NCH₂, CH₂Ar*), 2.26-2.18 (m, 2H, NCH₂, CH₂Ar*), 2.23 (s,3H, ArCH₂*), 2.18 (s, 3H, ArCH₃), 1.62-1.38 (m, 6H, HOCH₂CH₂CH₂CH₂). ¹³CNMR (100 MHz, CDCl₃), δ156.5, 150.7, 148.5, 145.3, 144.5, 144.3, 144.1,142.6, 141.4, 128.1, 127.8, 127.2 (2), 125.5, 125.3, 124.2, 123.6,123.0, 120.5, 120.1, 116.4, 114.5, 75.8, 66.9, 66.6, 64.5, 63.6, 63.0,61.4, 61.1, 61.0, 60.9 (2), 58.4, 58.2, 50.6, 47.4, 47.3, 33.3, 32.8,31.3, 29.9, 24.7, 21.7, 10.2, 9.8. FTIR (neat film), cm⁻¹ 3392 (m, NH),2939 (s), 2861 (m), 1702 (s, NCO₂), 1451 (m), 1416 (m), 1233 (m), 1112(m), 1052 (s), 1009 (m), 910 (m), 733 (s). R_(f) 0.25, 90% ethylacetate-hexanes. HRMS (TOF-ES⁺) Calcd for C₄₉H₆₁N₄O₁₀ (M+H)⁺: 865.4387,Found: 865.4390.

Substrate phenol resin (2.19 g, 355.5 μmol, 1.0 equiv, 0.1681 mmol/g)was suspended in 20.0 mL 20% (v/v) piperidine-N,N-dimethylformamide. Theresulting clear suspension of orange resin was stirred for 1.5 hr at 23°C. before removing the reaction supernatant via crimped cannula. Theproduct resin was sequentially washed with 3×20 mLN,N-dimethylformamide, 3×20 mL tetrahydrofuran, and 2×20 mLdichloromethane and dried in vacuo to provide a free-flowing orangeresin. Soluble substrate was isolated for characterization by incubatinga sample of product resin 12 (74.3 mg, 12.53 μmol, 1.0 equiv) in 2.0 mLdichloromethane with methanol (50.8 μL, 1.25 mmol, 100.0 equiv) andconcentrated hydrochloric acid (5.1 μL, 61.42 μmol, 4.9 equiv) for 18.5hr at 23° C. The reaction supernatant from this mixture was thencollected via crimped cannula along with 6×2 mL dichloromethane washesof the treated resin. The combined washes and supernatant werepartitioned between 30 mL dichloromethane and 30 mL sat. aq. sodiumbicarbonate and the organic layer isolated. After further extraction ofthe aqueous layer (2×20 mL diethyl ether), all organic extracts werecombined, dried over sodium sulfate, and concentrated in vacuo.Chromatographic purification of the resulting oil (SiO₂, 2%methanol-dichloromethane, column packed with eluent+1% triethylamine)afforded the expected cis-tetrahydroisoquinoline product as a whitesolid (1.6 mg, 20%).

cis-12: ¹H NMR (400 MHz, CDCl₃), * denotes non-fused aromatic ringprotons, δ6.78 (s, 1H, ArH), 6.66 (s, 1H, ArOH), 5.60 (s, 1H, ArOH*),3.98 (br s, 1H, NCHAr), 3.94 (d, 1H, J=10.8 Hz, ROCH₂CH₂), 3.82 (s, 3H,ArOCH₃*), 3.77 (s, 3H, ArOCH₃*), 3.73 (m, 1H, ROCH₂CH₂), 3.68 (s, 3H,ArOCH₃), 3.66 (s, 3H, ArOCH₃), 3.64 (t, 2H, J=6.0 Hz, HOCH₂), 3.54 (m,2H, OCHCH₂, CHCN), 3.32 (dd, 1H, J=4.4, 15.2 Hz, CH₂Ar), 3.17 (m, 1H,NCH₂), 2.92 (d, 1H, J=10.8 Hz, NCH₂), 2.79-2.71 (m, 3H, CHNCH₃, CHNH₂,NCH₂), 2.62 (s, 3H, NCH₃), 2.52 (dt, 1H, J=2.8, 10.8 Hz, NCH₂), 2.41(dd, 1H, J=13.2, 15.2 Hz, CH₂Ar), 2.24-2.20 (m, 2H, 2×CH₂Ar*), 2.22 (s,3H, ArCH₃*), 2.21 (s, 3H, ArCH₃), 1.60-1.38 (m, 6H, HOCH₂CH₂CH₂CH₂). ¹³CNMR (100 MHz, CDCl₃), δ150.4, 148.7, 146.4, 145.9, 145.0, 143.7, 124.8,124.1, 123.4, 122.3, 116.6, 114.8, 75.7, 66.5, 64.9, 62.9, 61.7, 61.4,61.1, 60.9, 60.8, 59.9, 59.7, 59.1, 58.2, 50.7, 47.3, 33.2, 32.7, 23.9,21.7, 10.2, 9.8. FTIR (neat film), cm⁻¹ 3354 (w, OH/NH), 2936 (m), 2859(w), 1455 (m), 1416 (m), 1112 (s), 1053 (m), 1005 (m). R_(f) 0.49, 10%methanol-dichloromethane, triethylamine-dipped plate. HRMS (TOF-ES⁺)Calcd for C₃₄H₅₁N₄O₈ (M+H)⁺: 643.3707, Found: 643.3713.

A solution of N-(9-fluorenylmethoxycarbonyl)-glycinal (Prepared asreported in Myers, A. G.; Kung, D. W. J. Am. Chem. Soc. 1999, 121,10828-10829) (25.8 mg, 91.50 μmol, 3.0 equiv) in 3.1 mL dichloroethanewas freeze-pump-thaw deoxygenated (3 cycles) and added via cannula toamine resin 12 (180.8 mg, 30.50 μmol, 1.0 equiv). The resulting resinsuspension was stirred at 40° C. for 20 hr before removing the reactionsupernatant via crimped cannula The product resin was then washed with6×3 mL tetrahydrofuran and 3×3 mL dichloromethane and dried in vacuo,yielding a free-flowing orange resin. Photometric determination of theproduct resin 9-fluorenylmethoxycarbonyl (Fmoc) loading level (A₂₉₀, 20%piperidine-N,N-dimethylformamide, duplicate measurements) established anFmoc-loading level of 0.2076 mmol/g, consistent with a yield of >100%for this transformation, suggesting that product resin 13 has theability to bind extra equivalents of aldehyde. An isolated yield forthis transformation was obtained by incubating product resin 13 (88.58mg, 14.31 μmol, 1.0 equiv) in 2.3 mL dichloromethane with methanol (57.9μL, 1.43 mmol, 100.0 equiv) and concentrated hydrochloric acid (4.8 μL,57.2 μmol, 4.9 equiv) for 19 hr at 23° C. The reaction supernatant wasthen collected via crimped cannula along with 6×2 mL dichloromethanewashes of the treated resin and the combined filtrates partitionedbetween 30 mL dichloromethane and 30 mL sat. aq. sodium bicarbonate. Theorganic layer was isolated and the aqueous layer further extracted with2×30 mL diethyl ether. All organic extracts were then combined, driedover sodium sulfate, and concentrated in vacuo to provide an orange oil.Chromatographic purification of this oil (prep TLC, SiO₂, 10%methanol-dichloromethane) afforded the expectedcis-/cis-bis-tetrahydroisoquinoline cleavage product (5.7 mg, 44%, 3steps) as a yellow-orange solid along with several diastereomericproducts (various centers, 5.8 mg, 45%, 3 steps) that were not cleanlyseparated by prep TLC. The combined 89% isolated yield over three stepscorresponds to an average yield per step of ˜96%.

cis-13: ¹H NMR (500 MHz, CDCl₃), * denotes unmethylatedtetrahydroisoquinoline ring system, δ7.76 (dd, 2H, J=3.5, 8.0 Hz, ArH),7.59-7.54 (m, 2H, ArH), 7.41-7.38 (m, 2H, ArH), 7.30 (q, 2H, J=7.0 Hz,ArH), 5.72 (br s, 1H, ArOH), 5.37 (br s, 1H, NHFmoc), 4.37 (d, 2H, J=7.5Hz, CO₂CH₂, NCHAr), 4.34 (d, 1H, CO₂CH₂), 4.21 (t, 1H, J=6.5 Hz,CO₂CH₂CH), 3.91 (d, 2H, J=11.5 Hz, ROCH₂CH₂, NCHAr*), 3.76 (s, 3H,ArOCH₃*), 3.74 (s, 3H, ArOCH₃*), 3.71 (m, 1H, ROCH₂CH₂), 3.68 (m, 1H,OCHCH₂), 3.66 (2×s, 6H, 2×ArOCH₃), 3.64 (s, 1H, CHCN), 3.49 (m, 4H,HOCH₂, CH₂NHFmoc), 3.27 (d, 1H, J=11.5 Hz, CH₂Ar), 2.84 (d, 2H, J=11.0Hz, NCH₂, CH₂Ar*), 2.71-2.68 (br s, 3H, NCH₃), 2.67-2.56 (m, 2H, CHNCH₃,CHNH), 2.48 (dt, 2H, J=3.5, 11.0 Hz, 2×NCH₂), 2.42 (m, 1H, CH₂Ar),2.24-2.21 (m, 2H, NCH₂, CH₂Ar*), 2.22 (s, 3H, ArCH₃*), 2.21 (s, 3H,ArCH₃), 1.61-1.54 (m, 2H, HOCH₂CH₂), 1.54-1.46 (m 2H, HOCH₂CH₂CH₂CH₂),1.46-1.34 (m, 2H, HOCH₂CH₂CH₂). ¹³C NMR (100 MHz, CDCl₃), δ157.5, 149.4,148.7, 145.7, 144.3, 143.9, 143.7, 142.0, 141.5, 131.1, 130.8, 128.6,127.9, 127.3, 126.2, 125.4, 123.7, 122.6, 120.2, 116.3, 75.8, 67.1,66.5, 64.9, 63.0, 61.4, 61.1, 61.0, 60.7, 59.1, 58.3, 52.8, 49.9, 47.4,47.0, 46.6, 33.3, 32.8, 29.9, 27.2, 24.2, 21.8, 9.9, 9.8. FTIR (neatfilm), cm⁻¹ 3390 (m, NH/OH), 2936 (s), 2863 (m), 1713 (m, NCO₂), 1451(s), 1413 (m), 1258 (m), 1112 (s), 1057 (s), 1006 (m), 734 (s). R_(f)0.34, 10% methanol-ichloromethane. HRMS (TOF-ES⁺) Calcd for C₅₁H₆₄N₅O₁₀(M+H)⁺: 906.4653, Found: 906.4688.

2,2,2-trifluoroethanol (35.2 μL, 483.10 μmol, 68.6 equiv), zinc chloride(0.5 M in tetrahydrofuran, 70.4 μL, 35.21 μmol, 5.0 equiv), andtrimethylsilylcyanide (3.8 μL, 28.17 μmol, 4.0 equiv) were sequentiallyadded to a suspension of resin 13 (43.6 mg, 7.04 μmol, 1.0 equiv) in105.6 μL tetrahydrofuran at 23° C. The resulting yellow suspension wasstirred for 16 hr at 23° C. before diluting the reaction mixture with1.5 mL tetrahydrofuran. The reaction supernatant was then collected viacrimped cannula along with 5×2 mL tetrahydrofuran washes of the treatedresin. The collected filtrates were partitioned between 5 mL ethylacetate and 15 mL of an aqueous solution of 0.2 Nethylenediaminetetraacetic acid disodium salt dihydrate and 0.4 N sodiumhydroxide (pH 10). The organic layer was isolated and washed with 1×10mL brine and the aqueous layers then combined and extracted with afurther 2×15 mL ethyl acetate. The combined organic extracts were thendried over sodium sulfate and concentrated in vacuo. Chromatographicpurification of the resulting red oil (prep TLC, SiO₂, 5%methanol-dichloromethane) afforded pentacyclic product 2a as a whitesolid (2.0 mg, 41%) along with small amounts of the product aminonitrilehydrolysis product (0.4 mg, 8%).

2a: ¹H NMR (400 MHz, CDCl₃), δ7.76 (t, 2H, J=6.8 Hz, ArH), 7.47-7.39 (m,2H, ArH), 7.30 (t, 2H, J=7.2 Hz, ArH), 5.57 (br s, 1H, ArOH), 5.51 (brs, 1H, ArOH), 4.53 (t, 1H, J=6.0 Hz, NHFmoc), 4.32 (dd, 1H, J=6.8, 10.8Hz, CO₂CH₂), 4.22 (dd, 1H, J=6.0, 10.8 Hz, CO₂CH₂), 4.14 (br s, 1H,ArCHNCH₃), 4.11 (t, 1H, J=4.4 Hz, ArCHCH₂NHFmoc), 4.06 (t, 1H, J=6.4 Hz,CO₂CH₂CH), 3.74 (s, 3H, ArOCH₃), 3.60 (s, 6H, 2×ArOCH₂), 3.55 (s, 3H,ArOCH₃), 3.29 (d, 1H, J=7.2, ArCH₂CHNCH₃), 3.24-3.19 (m, 3H,ArCH₂CHNC(CN), ArCH₂CHNC(CN), CH₂NHFmoc), 3.11-3.04 (m, 1H, CH₂NHFmoc),2.96 (dd, 1H, J=8.0, 18.8 Hz, ArCH₂CHNCH₃), 2.33 (d, 1H, J=11.6 Hz,ArCH₂CHNCH₃), 2.31 (s, 3H, NCH₃), 2.19 (s, 3H, ArCH₃), 2.11 (s, 3H,ArCH₃), 1.88 (dd, 1H, J=11.6, 16.4 Hz, ArCH₂CHNC(CN)). R_(f) 0.38, 5%methanoldichloromethane. LRMS (TOF-ES⁺) Calcd for C₄₃H₄₇N₄O₈ (M+H)⁺:747, Found: 747.Exemplary Solid-Supported Syntheses of Compounds Having AlternatePentacyclic Core Structures:

A solution of hexanal (7.8 μL, 64.70 μmol, 5.0 equiv) in 1.3 mLdichloroethane was freeze-pump-thaw deoxygenated (3 cycles) and addedvia cannula to amine resin 12 (76.7 mg, 12.94 μmol, 1.0 equiv). Theresulting resin suspension was stirred in the dark at 40° C. for 20.5 hrbefore removing the reaction supernatant via crimped cannula. Theproduct resin was then washed with 6×3 mL tetrahydrofuran and 3×3 mLdichloromethane and dried in vacuo, yielding a free-flowing orangeresin. The product resin was directly employed in subsequent reactions.

Zinc chloride (0.5 M in tetrahydrofuran, 2.3 mL, 1.16 mmol, 100.0 equiv)and trimethylsilylcyanide (151.8 μL, 1.14 mmol, 99.0 equiv) weresequentially added to a suspension of substrate resin (69.8 mg, 11.61μmol, 1.0 equiv) in 460.0 μL 2,2,2-trifluoroethanol at 23° C. Theresulting yellow suspension was stirred for 15.5 hr at 23° C. beforeremoving and collecting the reaction supernatant via crimped cannulaalong with 5×3 mL tetrahydrofuran washes of the treated resin. Thecollected filtrates were partitioned between 10 mL ethyl acetate and 20mL of an aqueous solution of 0.2 N ethylenediaminetetraacetic aciddisodium salt dihydrate and 0.4 N sodium hydroxide (pH 10). The organiclayer was isolated and washed with 1×15 mL brine and the aqueous layersthen combined and extracted with a further 2×15 mL ethyl acetate. Thecombined organic extracts were then dried over sodium sulfate andconcentrated in vacuo. Chromatographic purification of the resulting redoil (SiO₂, 40% ethyl acetate-hexanes) afforded the pentacyclic productas a white solid (1.9 mg, 29%, 4 steps).

¹H NMR (500 MHz, C₆D₆), ˜2:1 mixture of rotamers, * denotes minorrotamer signals, δ 5.75 (s, 1H, ArOH), 5.18 (s, 1H, ArOH*), 5.17 (s, 1H,ArOH), 4.20 (m, 1H, ArCHNCH₃), 4.16 (d, 1H, J=2.0 Hz, ArCHNCH₃*), 3.93(m, 1H, ArCHNC(CN)*), 3.90 (m, 1H, ArCHNC(CN)), 3.83 (d, 1H, J=3.2 Hz,CHCN), 3.73 (d, 1H, CHCN*), 3.53 (s, 3H, ArOCH₃*), 3.38 (s, 3H, ArOCH₃),3.31 (s, 3H, ArOCH₃), 3.22 (s, 3H, ArOCH₃*), 3.14 (s, 3H, ArOCH₃*), 3.08(s, 3H, ArOCH₃), 3.02 (s, 3H, ArOCH₃), 3.00 (s, 3H, ArOCH₃*), 2.98 (m,1H, ArCH₂CHNCH₃), 2.80 (d, 1H, J=13.4 Hz, ArCH₂CHNC(CN)), 2.66 (dd, 1H,J=8.0, 18.8 Hz, ArCH₂CHNC(CN)), 2.58 (dt, 1H, J=4.4, 12.8 Hz,ArCH₂CHNCH₃), 2.23 (dt, 1H, 4.4, 18.0 Hz, ArCH₂CHNCH₃), 2.15 (s, 3H,NCH₃), 2.13 (m, 1H, ArCH₂CHNC(CN)), 2.12 (s, 3H, ArCH₃*), 2.11 (s, 3H,ArCH₃), 2.05 (s, 3H, ArCH₃), 2.04 (m, 1H, ArCHCH₂), 1.95 (d, 1H, J=18.8Hz, ArCHCH₂), 1.62 (dt, 2H, J=4.8, 13.2 Hz, ArCHCH₂CH₂), 0.90-0.78 (m,4H, CH₂CH₂CH₂CH₃), 0.65 (t, 3H, J=7.2 Hz, CH₂CH₃). FTIR (neat film),cm⁻¹ 2934 (m), 2859 (w), 1462 (s), 1414 (s), 1280 (m), 1109 (m), 1063(m), 1005 (m). R_(f) 0.32, 5% methanol-dichloromethane.

A solution of ethyl glyoxylate (50% w/w in toluene, 13.0 μL, 65.79 μmol,5.0 equiv) in 1.3 mL dichloroethane was freeze-pump-thaw deoxygenated (3cycles) and added via cannula to amine resin 12 (78.0 mg, 13.16 μmol,1.0 equiv). The resulting resin suspension was stirred in the dark at23° C. for 22 hr before removing the reaction supernatant via crimpedcannula. The product resin was then washed with 2×3N,N-dimethylformamide, 4×3 mL tetrahydrofuran, and 2×2 mLdichloromethane and dried in vacuo, yielding a free-flowing orangeresin. The product resin was directly employed in subsequent reactions.

2,2,2-trifluoroethanol (61.6 μL, 845.4 μmol, 68.6 equiv), zinc chloride(0.5 M in tetrahydrofuran, 123.3 μL, 61.64 μmol, 5.0 equiv), andtrimethylsilylcyanide (6.6 μL, 49.31 μmol, 4.0 equiv) were sequentiallyadded to a suspension of substrate resin (74.1 mg, 12.33 μmol, 1.0equiv) in 185.0 μL tetrahydrofuran at 23° C. The resulting yellowsuspension was stirred for 7 hr at 23° C. before diluting the reactionmixture with 1.5 mL tetrahydrofuran. The reaction supernatant was thencollected via crimped cannula along with 5×2 mL tetrahydrofuran washesof the treated resin. The collected filtrates were partitioned between 5mL ethyl acetate and 15 mL of an aqueous solution of 0.2 Nethylenediaminetetraacetic acid disodium salt dihydrate and 0.4 N sodiumhydroxide (pH 10). The organic layer was isolated and washed with 1×10mL brine and the aqueous layers then combined and extracted with afurther 2×15 mL ethyl acetate. The combined organic extracts were thendried over sodium sulfate and concentrated in vacuo. Chromatographicpurification of the resulting orange oil (prep TLC, SiO₂, 5%methanol-dichloromethane) afforded the pentacyclic product as ayellowish-white solid (2.2 mg, 31.5%, 4 steps)

¹H NMR (400 MHz, CDCl₃), δ5.56 (s, 1H, ArOH), 5.51 (s, 1H, ArOH), 4.57(s, 1H, ArCHCO₂), 4.35 (d, 1H, J=2.4 Hz, CHCN), 4.17 (dd, 1H, J=1.2, 2.8Hz, ArCHNCH₃), 4.04-3.97 (m, 1H, CO₂CH₂), 3.95-3.85 (m, 1H, CO₂CH₂),3.75 (s, 3H, ArOCH₃), 3.74 (s, 3H, ArOCH₃), 3.65 (s, 3H, ArOCH₃), 3.63(s, 3H, ArOCH₃), 3.39 (d, 1H, J=8.0 Hz, ArCH₂CHC(CN)), 3.26 (dt, 1H,J=2.4, 11.2 Hz, ArCH₂CHNC(CN)), 3.22 (dd, 1H, J=2.8, 15.6 Hz,ArCH₂CHNC(CN)), 3.03 (dd, 1H, J=8.0, 18 Hz, ArCH₂CHNCH₃), 2.41 (d, 1H,J=18.4 Hz, ArCH₂CHNCH₃), 2.30 (s, 3H, NCH₃), 2.21 (s, 3H, ArCH₃), 2.20(s, 3H, ArCH₃), 2.03 (dd, 1H, J=11.6, 15.2 Hz, ArCH₂CHNC(CN)), 1.01 (t,3H, J=7.2 Hz, CO₂CH₂CH₃). FTIR (neat film), cm⁻¹ 3422 (m, OH), 2936 (m),2829 (w), 1728 (m, CO₂), 1464 (s), 1414 (s), 1276 (m), 1152 (m), 1061(s), 1027 (m), 1004 (m). R_(f) 0.39, 5% methanol-dichloromethane. HRMS(TOF-ES⁺) Calcd for C₃₀H₃₈N₃O₈ (M+H)⁺: 568.2658, Found: 568.2677.

A solution of trans-cinnamaldehyde (8.8 μL, 69.42 μmol, 5.0 equiv) in1.35 mL dichloroethane was freeze-pump-thaw deoxygenated (3 cycles) andadded via cannula to amine resin 12 (82.3 mg, 13.88 μmol, 1.0 equiv).The resulting resin suspension was stirred in the dark at 40° C. for 21hr before removing the reaction supernatant via crimped cannula. Theproduct resin was then washed with 2×3 N,N-dimethylformamide, 4×3 mLtetrahydrofuran, and 2×2 mL dichloromethane and dried in vacuo, yieldinga free-flowing orange resin. The product resin was directly employed insubsequent reactions.

2,2,2-trifluoroethanol (65.0 μL, 892.1 μmol, 68.3 equiv), zinc chloride(0.5 M in tetrahydrofuran, 130.6 μL, 65.29 μmol, 5.0 equiv), andtrimethylsilylcyanide (7.0 μL, 52.23 μmol, 4.0 equiv) were sequentiallyadded to a suspension of substrate resin (78.9 mg, 13.06 μmol, 1.0equiv) in 196.0 μL tetrahydrofuran at 23° C. The resulting yellowsuspension was stirred for 5.5 hr at 23° C. before diluting the reactionmixture with 1.5 mL tetrahydrofuran. The reaction supernatant was thencollected via crimped cannula along with 5×2 mL tetrahydrofuran washesof the treated resin. The collected filtrates were partitioned between 5mL ethyl acetate and 15 mL of an aqueous solution of 0.2 Nethylenediaminetetraacetic acid disodium salt dihydrate and 0.4 N sodiumhydroxide (pH 10). The organic layer was isolated and washed with 1×10mL brine and the aqueous layers then combined and extracted with afurther 2×15 mL ethyl acetate. The combined organic extracts were thendried over sodium sulfate and concentrated in vacuo. Chromatographicpurification of the resulting yellow-brown oil (prep TLC, SiO₂, 5%methanol-dichloromethane) afforded the pentacyclic product as a whitesolid (0.3 mg, 4%, 4 steps)

¹H NMR (400 MHz, CDCl₃), δ7.21-7.12 (m, 5H, ArH), 6.42 (d, 1H, J=15.2Hz, CHCHAr), 6.03 (dd, 1H, J=5.6, 15.6 Hz, CHCHAr), 5.56 (s, 1H, ArOH),5.49 (s, 1H, ArOH), 4.59 (d, 1H, J=6.0 Hz, ArCHCHCHAr), 4.21 (d, 1H,J=2.4 Hz, CHCN), 4.02 (d, 1H, J=2.8 Hz, ArCHNCH₃), 3.78 (s, 3H, ArOCH₃),3.73 (s, 3H, ArOCH₃), 3.61 (s, 3H, ArOCH₃), 3.60 (s, 3H, ArOCH₃), 3.39(d, 1H, J=6.4 Hz, ArCH₂CHC(CN)), 3.32 (dt, 1H, J=12.0 Hz,ArCH₂CHNC(CN)), 3.26 (dd, 1H, J=2.4, 15.6 Hz, ArCH₂CHNC(CN)), 3.04 (dd,1H, J=8.0, 18.4 Hz, ArCH₂CHNCH₃), 2.48 (d, 1H, J=18.8 Hz, ArCH₂CHNCH₃),2.35 (s, 3H, NCH₃), 2.28 (s, 3H, ArCH₃), 2.18 (s, 3H, ArCH₃), 2.00 (dd,1H, J=12.8, 14.0 Hz, ArCH₂CHNC(CN). FTIR (neat film), cm⁻¹ 3357 (w, OH),2918 (m), 2849 (w), 1463 (s), 1415 (s), 1109 (s), 1060 (s), 1002 (s),744 (m). R_(f) 0.37, 5% methanol-chloromethane. HRMS (ApCI⁺) Calcd forC₃₄H₃₉N₂O₆ (M-CN)⁺: 571, Found: 571.

A solution of benzaldehyde boronate ester (5.2 mg, 23.87 μmol, 5.0equiv) in 600 μL 1,2-dichloroethane was freeze-pump-thaw deoxygenated (3cycles) and then added via cannula to the amine resin (28.3 mg, 4.77μmol, 0.1687 mmol/g, 1.0 equiv). The resulting suspension was stirred at75° C. for 31 h and the supernatant was then removed via cannula. Theproduct resin was washed with 4×1 mL N,N-dimethylformamide, 6×1 mLtetrahydrofuran, and 2×1 mL diethyl ether and then dried in vacuo toprovide 26.2 mg free-flowing orange resin.

-   R_(f) 0.48 (methanolysis product, decomposes), 10%    methanoldichloromethane.

Zinc chloride (25.7 μL, 0.5 M in tetrahydrofuran, 12.83 μmol, 3.0 equiv)was added to a mixture of boronate resin (26.2 mg, 4.28 μmol, 0.1632mmol/g, 1.0 equiv) and 4 Å molecular sieves (2.5 mg) suspended in 160.0μL tetrahydrofuran. The resulting suspension was stirred at 55° C. for1.5 h. The reaction supernatant was then removed via crimped cannula andcollected along with 5×1.5 mL dichloromethane washes of the productresin. The combined superntant and washes were partitioned between 15 mLdichloromethane and 15 mL aqueous phosphate buffer solution (0.05 Msodium phosphate monobasic, 0.05 M potassium phosphate dibasic, pH 7)and the organic layer isolated. The organic layer was then washed withan additional 8 mL phosphate buffer solution, dried over sodium sulfate,and concentrated in vacuo to provide an orangish-brown oil.Lyophilization of this oil from 300 μL benzene afforded the heptacyclicproduct as a tan solid (2.2 mg, 57.7% yield (9 steps)).

¹H NMR (600 MHz, CDCl₃), δ7.53 (d, 2H, J=7.8 Hz, ArH), 7.16 (d, 2H,J=7.8 Hz, ArH), 5.58 (s, 1H, ArOH), 5.30 (s, 1H, ArOH), 4.89 (s, 1H,CHAr₁Ar₂), 4.23 (d, 1H, J=3.0 Hz, N(CH₃)CH), 3.80 (s, 3H, ArOCH₃), 3.73(s (obsc.), 1H, CHCN), 3.71 (s, 6H, 2×ArOCH₃), 3.64 (s, 2H, BOCH₂), 3.63(s, 2H, BOCH₂), 3.47 (s, 3H, ArOCH₃), 3.39 (dt, 1H, J=10.8 Hz,N(CH₃)CHCH), 3.31 (dd, 1H, J=2.4, 16.2 Hz, N(CH₃)CHCHCH₂), 3.25 (d, 1H,J=8.4 Hz, NCH(CN)CH), 2.83 (dd, 1H, J=8.4, 18.0 Hz, NCH(CN)CHCH₂), 2.26(s, 3H, NCH₃), 2.25 (s, 3H, ArCH₃), 2.18 (dd, 1H, J=5.4, 15.6 Hz,N(CH₃)CHCHCH₂), 2.15 (s, 3H, ArCH₃), 1.84 (d, 1H, J=18.0 Hz,NCH(CN)CHCH₂), 0.98 (s, 6H, C(CH₂)₂). R_(f) 0.27 (decomposes), 5%methanol-dichloromethane. HRMS (TOF-ES⁺) Calcd for C₃₈H₄₇BN₃O₈ ⁺ (M+H)⁺:684.3456, Found: 684.3487.

Exemplary Alternate N-alkylation Reaction:

N,N-diethylaniline (34.9 μL, 219.0 μmol, 45.0 equiv) was added to asuspension of amine resin (23.5 mg, 4.87 μmol, 0.2071 mmol/g, 1.0 equiv)in 390.0 μL N,N-diethylformamide. Iodoacetonitrile (14.1 μL, 194.67μmol, 40.0 equiv) was then added and the resulting suspension stirredfor 4 h at 55° C. The reaction supernatant was then removed via acrimped cannula and the product resin washed with 4×2 mLN,N-dimethylformamide, 4×2 mL tetrahydrofuran, and 2×2 mL diethyl ether.The product resin was then dried in vacuo to provide a free-flowingred-brown resin.

-   R_(f) 0.41, 0.45 (methanolysis products, diastereomers), 10%    methanol-dichloromethane.

Acetic acid (1.1 μL, 18.65 μmol, 4.0 equiv) and tetrabutylammoniumfluoride (1.0 M in tetrahydrofuran, 9.3 μL, 9.30 μmol, 2.0 equiv) weresequentially added to a suspension of silylphenol resin (22.7 mg, 4.66μmol, 0.2054 mmol/g, 1.0 equiv) in 230 μL tetrahydrofuran. The resultingsuspension was stirred at 23° C. for 2.5 h before removal of thesuperntant solution via crimped cannula. Washing of the product resinwith with 4×1 mL N,N-dimethylformamide, 4×1 mL tetrahydrofuran, and 2×1mL diethyl ether and drying in vacuo afforded the phenol resin as afree-flowing brown-orange resin.

Piperidine (60.0 μL, 620.57 μmol, 133 equiv) was added to a suspensionof phenol resin (22.2 μγ, 4.66 μmol, 1.0 equiv) in 240 μLN,N-diethylformamide and the resulting suspension was stirred for 3.5 hat 23° C. The supernantant solution was then removed via crimped cannulaand the product resin washed with 4×1 mL N,N-dimethylformamide, 6×1 mLtetrahydrofuran, and 2×1 mL diethyl ether. Drying of the washed resin invacuo provided the immbolized aminophenol as a free-flowing orangeresin.

A solution of N-Fmoc glycinal (3.9 mg, 13.98 μmol, 3.0 equiv) in 350 μL1,2-dichloroethane was freeze-pump-thaw deoxygenated (3 cycles) and thenadded via cannula to the aminophenol resin (21.2 mg, 4.66 μmol, 1.0equiv). The resulting suspension was stirred at 55° C. for 17 h beforethe supernatant solution was removed via crimped cannula. The productresin was then washed with 3×1 mL N,N-dimethylformamide, 5×1 mLtetrahydrofuran, and 2×1 mL diethyl ether and dried in vacuo to affordthe immbolilized bistetrahydroisoquinoline as a free-flowingbrown-orange resin.

-   R_(f) 0.47 (methanolysis product), 10% methanolchloromethane.

Zinc chloride (29.0 μL, 0.5 M in tetrahydrofuran, 14.51 μmol, 3.0 equiv)was added to a mixture of N-Fmoc resin (23.2 mg, 4.84 μmol, 0.2085mmol/g, 1.0 equiv) and 4 Å molecular sieves (2.5 mg) suspended in 180.0μL tetrahydrofuran. The resulting suspension was stirred at 55° C. for1.5 h before being diluted with 650 μL dichloromethane. The resultingsuspension was loaded onto a short silica gel plug and eluted with 4plug-volumes of 10% tetrahydrofuran-dichloromethane. The eluted solventwas concentrated in vacuo to afford the product bis-aminonitrile as anoff-white solid (0.8 mg, 21.4% yield (9 steps)).

¹H NMR (600 MHz, CDCl₃), δ7.78 (m, 2H, J=7.8 Hz, ArH), 7.60 (d, 1H,J=7.8 Hz, ArH), 7.49 (m, 1H, ArH), 7.41 (m, 2H, ArH), 7.32 (m, 2H, ArH),5.56 (s, 1H, ArOH), 5.54 (s, 1H, ArOH), 4.47 (t, 1H, NHFmoc), 4.31 (t,1H, J=6.0 Hz, CO₂CH₂), 4.29 (s, 1H, N(CH₂CN)CHAr), 4.24 (t, 1H, J=7.8Hz, CO₂CH₂), 4.11 (t, 1H, J=4.2 Hz, NCHCH₂NHFmoc), 4.07 (t, 1H,CO₂CH₂CH), 3.75 (s, 3H, ArOCH₃), 3.73 (s, 3H, ArOCH₃), 3.63 (s, 1H,NCH(CN)CH), 3.61 (s, 3H, ArOCH₃), 3.48 (s, 3H, ArOCH₃), 3.40 (d, 1H,J=16.8 Hz, NCH₂CN), 3.34 (d, 1H, J=16.8 Hz, NCH₂CN), 3.28-3.17 (m, 3H,CH₂NHFmoc, N(CH₂CN)CHCH, N(CH₂CN)CHCHCH₂), 2.93-2.85 (m, 2H, CH₂NHFmoc,NCH(CN)CHCH₂), 2.36 (d, 1H, J=19.2 Hz, NCH(CN)CHCH₂), 2.19 (s, 3H,ArCH₃), 2.17 (s, 3H, ArCH₃), 1.85 (m, 1H, N(CH₂CN)CHCHCH₂). FTIR (neatfilm), cm⁻¹ 3350 (m, OH), 2925 (m), 1714 (s), 1455 (m), 1415 (m), 1109(s), 1059 (s), 760 (m). R_(f) 0.50, 5% methanol-dichloromethane. HRMS(TOF-ES⁺) Calcd for C₄₄H₄₆N₅O₈ ⁺ (M+H)⁺: 772.3346, Found: 772.3369.

4) Synthesis and Characterization of Certain Exemplary Analogues:

It will be appreciated that although the synthesis of certain of thecompounds are described using traditional solution phase techniques (forexample as described in section 1 and 2 above) and the synthesis ofcertain of the compounds are described using solid-supported techniques(for example, as described in section 3 above), each of the compoundsdescribed below can be prepared using either traditional solution phasetechniques or solid-supported techniques.

EXAMPLE 1 bis-2-Pyridyl Derivative

Pyridine 2-carboxaldehyde (0.29 μL, 3.0 μmol, 2.0 equiv) and sodiumtriacetoxy-borohydride (0.48 mg, 2.3 μmol, 1.5 equiv) were addedsequentially, each in one portion, to a stirred solution of the amine(0.8 mg, 1.5 μmol, 1 equiv) in acetonitrile (0.1 mL) at 24° C. under anargon atmosphere. The mixture was stirred for 1 h and then a secondportion of pyridine 2-carboxaldehyde (0.29 μL, 3.0 μmol, 2.0 equiv) andsodium triacetoxyborohydride (0.48 mg, 2.3 μmol, 1.5 equiv) were addedsequentially, each in one portion. The mixture was stirred for a further30 min, then was diluted with ethyl acetate (10 mL) and washed with a1:1 mixture of brine solution and saturated aqueous sodium hydrogencarbonate solution (2×3 mL). The organic layer was dried over sodiumsulfate and concentrated in vacuo to leave a colorless oil. Purificationby flash column chromatography (ethyl acetate→80% ethylacetate-methanol) gave the bis-2-pyridyl derivative (0.8 mg, 85%) as awhite solid.

¹H NMR (500 MHz, CDCl₃), δ10.47 (br. s, 1H, NH), 8.51 (d, 1H, J=4.9,ArH), 7.64 (ddd, 1H, J=9.4, 7.8, 1.7, ArH), 7.37 (d, 1H, J=7.8, ArH),7.19 (m, 1H, ArH), 5.48 (s, 1H, ArOH), 4.27 (d, 1H, J=2.5, CHC≡N), 4.10(br. d, 1H, J=˜1.5, ArCHNCH₃), 4.00 (dd, 1H, J=9.3, 2.5, ArCHCH₂NH),3.82 (s, 3H, ArOCH₃), 3.81 (s, 4H, NCH₂Ar), 3.75 (s, 3H, ArOCH₃), 3.62(s, 3H, ArOCH₃), 3.58 (s, 3H, ArOCH₃), 3.38 (br. d, 1H, J=˜9.8,CHCHC≡N), 3.19-3.12 (m, 2H, ArCHCHCH₂Ar, ArCHCHCH₂Ar), 3.05 (dd, 1H,J=18.1, 8.3, CH₂CHCHC≡N), 2.88 (dd, 1H, J=13.7, 2.4, ArCHCH₂N), 2.62(dd, 1H, J=13.7, 9.3, ArCHCH₂N), 2.45 (d, 1H, J=18.1, CH₂CHCHC≡N), 2.28(s, 3H, NCH₃), 2.23 (s, 3H, ArCH₃), 2.18 (s, 3H, ArCH₃), 1.73 (dd, 1H,J=14.9, 11.0, ArCHCHCH₂). FTIR (neat film), cm⁻¹ 3354, 2923, 2687, 2226,1456. HRMS (ES) Calcd for C₄₀H₄₇N₆O₆ (MH)⁺: 707.3557, Found: 707.3580.

EXAMPLE 2 2-Furylmethyl Derivative

Furaldehyde (0.32 μL, 3.8 μmol, 2.0 equiv) and sodiumtriacetoxyborohydride (0.61 mg, 2.9 μmol, 1.5 equiv) were addedsequentially, each in one portion, to a stirred solution of the amine(1.0 mg, 1.9 μmol, 1 equiv) in acetonitrile (0.1 mL) at 24° C. under anargon atmosphere. The mixture was stirred for 50 min, then was dilutedwith ethyl acetate (10 mL) and washed with a 1:1 mixture of brinesolution and saturated aqueous sodium hydrogen carbonate solution (2×3mL). The organic layer was dried over sodium sulfate and concentrated invacuo to leave a brown oil. Purification by flash column chromatography(80% ethyl acetate-hexanes) gave the 2-furylmethyl derivative (1.1 mg,95%) as a white solid.

¹H NMR (500 MHz, CDCl₃), δ7.34 (dd, 1H, J=1.0, 0.9, ArH), 6.30 (dd, 1H,J=3.2, 1.7, ArH), 6.10 (app. d, 1H, J=˜2.4, ArH), 5.52 (s, 1H, ArOH),4.15 (dd, 1H, J=2.9, 1.0, ArCHNCH₃), 4.00 (dd, 1H, J=7.9, 3.0,ArCHCH₂NH), 3.98 (d, 1H, J=2.5, CHC≡N), 3.79 (s, 3H, ArOCH₃), 3.76 (s,3H, ArOCH₃), 3.72 (qu, 2H, AB system, ArCH₂NH), 3.61 (s, 3H, ArOCH₃),3.60 (s, 3H, ArOCH₃), 3.38 (br. d, 1H, J=˜9.8, CHCHC≡N), 3.25-3.21 (m,2H, ArCHCHCH₂Ar, ArCHCHCH₂Ar), 3.05 (dd, 1H, J=18.5, 8.0, CH₂CHCHC≡N),2.66 (dd, 1H, J=13.2, 3.4, ArCHCH₂NH), 2.60 (dd, 1H, J=13.2, 8.1,ArCHCH₂NH), 2.36 (d, 1H, J=18.5, CH₂CHCHC≡N), 2.31 (s, 3H, NCH₃), 2.22(s, 3H, ArCH₃), 2.19 (s, 3H, ArCH₃), 1.82 (dd, 1H, J=15.6, 12.2,ArCHCHCH₂Ar). FTIR (neat film), cm⁻¹ 3395, 3313, 2933, 2226, 1462. HRMS(ES) Calcd for C₃₃H₄₁N₄O₇ (MH)⁺: 605.2975, Found: 605.2956.

EXAMPLE 3 L-Tryptophan Derivative

Palladium on carbon (10 wt %, 0.1 8 mg, 0.17 μmol, 0.1 equiv) was addedin one portion to the carbobenzyloxy-protected tryptophan derivative(1.5 mg, 1.7 μmol, 1 equiv) in methanol (0.5 mL) at 23° C. A hydrogenatmosphere was introduced by alternatively evacuating the reaction flaskand flushing with hydrogen. The reaction mixture was stirred for 3 h 20min, then was filtered through a short pad of Celite. Concentration invacuo left a white solid, which was purified by flash columnchromatography (ethyl acetate→10:1 ethyl acetate-methanol) to give theL-tryptophan derivative (0.5 mg, 42%) as a white solid along withrecovered starting material (0.5 mg, 33%).

¹H NMR (500 MHz, CDCl₃), δ8.17 (br. s, 1H, ArNH), 7.56 (d, 1H, J=7.8,ArH), 7.37 (d, 1H, J=8.3, ArH), 7.20 (m, 1H, ArH), 7.11 (m, 1H, ArH),6.84 (t, 1H, J=5.6, NHCO), 6.69 (d, 1H, J=2.4, ArH), 5.40 (s, 1H, ArOH),5.31 (s, 1H, ArOH), 4.14-4.10 (m, 2H, CHCH₂NHCO, ArCHNCH₃), 3.85 (d, 1H,J=2.4, CHC≡N), 3.76 (s, 3H, ArOCH₃), 3.70 (s, 3H, ArOCH₃), 3.61 (s, 3H,ArOCH₃), 3.66 (ddd, 1H, J=13.6, 6.8, 3.3, CH₂NHCO), 3.49 (s, 3H,ArOCH₃), 3.34-3.27 (m, 1H (CHCHC≡N), 1H (CH₂NHCO), 1H (NHCOCHNH₂)), 3.23(ddd, 1H, J=11.7, 2.4, 2.4, ArCHCHCH₂Ar), 3.16 (dd, 1H, J=9.0, 2.4,ArCHCHCH₂Ar), 3.02 (dd, 1H, J=14.6, 4.8, NHCOCHCH₂), 2.95 (dd, 1H,J=18.6, 7.9, CH₂CHCHC≡N), 2.69 (dd, 1H, J=14.6, 7.3, NHCOCHCH₂), 2.41(d, 1H, J=18.6, CH₂CHCHC≡N), 2.28 (s, 3H, NCH₃), 2.21 (s, 3H, ArCH₃),2.08 (s, 3H, ArCH₃), 1.89 (dd, 1H, J=15.0, 10.7, ArCHCHCH₂). FTIR (neatfilm), cm⁻¹ 3354, 2913, 2226, 1651, 1262. HRMS (ES) Calcd for C₃₉H₄₆N₆O₇(MH)⁺: 711.3506, Found: 711.3488.

EXAMPLE 4 Benzyl Derivative

Benzaldehyde (0.43 μL, 4.2 μmol, 2.0 equiv) and sodiumtriacetoxyborohydride (0.67 mg, 3.1 μmol, 1.5 equiv) were addedsequentially, each in one portion, to a stirred solution of the amine(1.0 mg, 1.9 μmol, 1 equiv) in acetonitrile (0.1 mL) at 23° C. under anargon atmosphere. The mixture was stirred for 45 min, then was dilutedwith ethyl acetate (10 mL) and washed with a 1:1 mixture of brinesolution and saturated aqueous sodium hydrogen carbonate solution (2×3mL). The organic layer was dried over sodium sulfate and concentrated invacuo to leave a colorless oil. Purification by flash columnchromatography (ethyl acetate) gave the benzyl derivative (1.2 mg, 93%)as a white solid.

¹H NMR (500 MHz, CDCl₃), δ7.33-7.28 (m, 3H, ArH), 7.19-7.17 (m, 2H,ArH), 5.52 (s, 1H, ArOH), 5.31 (s, 1H, ArOH), 4.14 (dd, 1H, J=2.9, 1.0,ArCHNCH₃), 4.01 (t, 1H, J=5.6, ArCHCH₂NH), 3.86 (d, 1H, J=2.4, CHC≡N),3.80 (s, 3H, ArOCH₃), 3.75 (s, 3H, ArOCH₃), 3.73 (d, 2H, J=3.0,ArCH₂NH), 3.61 (s, 3H, ArOCH₃), 3.56 (s, 3H, ArOCH₃), 3.35 (br. d, 1H,J=˜7.8, CHCHC≡N), 3.25-3.21 (m, 2H, ArCHCHCH₂Ar, ArCHCHCH₂Ar), 3.02 (dd,1H, J=18.6, 7.8, CH₂CHCHC≡N), 2.66 (d, 1H, J=5.3, ArCHCH₂NH), 2.33 (d,1H, J=18.6, CH₂CHCHC≡N), 2.30 (s, 3H, NCH₃), 2.20 (s, 6H, 2∞ArCH₃), 1.83(dd, 1H, J=15.8, 12.0, ArCHCHCH₂Ar). FTIR (neat film), cm⁻¹ 3395, 2923,2574, 2226, 1456, 1108. HRMS (ES) Calcd for C₃₅H₄₃N₄O₆ (MH)⁺: 615.3182,Found: 615.3176.

EXAMPLE 5 2-Pyridyl Derivative

Pyridine 2-carboxaldehyde (0.36 μL, 3.8 μmol, 2.0 equiv) and sodiumtriacetoxy-borohydride (0.61 mg, 2.9 μmol, 1.5 equiv) were addedsequentially, each in one portion, to a stirred solution of the amine(1.0 mg, 1.9 μmol, 1 equiv) in acetonitrile (0.15 mL) at 24° C. under anargon atmosphere. The mixture was stirred for 40 min, then was dilutedwith ethyl acetate (10 mL) and washed with a 1:1 mixture of brinesolution and saturated aqueous sodium hydrogen carbonate solution (2×3mL). The organic layer was dried over sodium sulfate and concentrated invacuo to leave a white solid. Purification by flash columnchromatography (ethyl acetate→90% ethyl acetate-methanol) gave the2-pyridyl derivative (0.6 mg, 51%) as a white solid.

¹H NMR (500 MHz, CDCl₃), δ8.53-8.52 (m, 1H, ArH), 7.63 (ddd, 1H, J=7.8,7.8, 2.0, ArH), 7.19 (ddd, 1H, J=7.8, 5.8, 1.0, ArH), 7.14 (d, 1H,J=7.8, ArH), 5.51 (s, 1H, ArOH), 4.15 (dd, 1H, J=2.5, 1.0, ArCHNCH₃),4.07 (d, 1H, J=2.4, CHC≡N), 4.05 (dd, 1H, J=8.3, 2.5, ArCHCH₂NH), 3.86(q, 2H, AB system, ArCH₂NH), 3.78 (s, 3H, ArOCH₃), 3.75 (s, 3H, ArOCH₃),3.61 (s, 3H, ArOCH₃), 3.52 (s, 3H, ArOCH₃), 3.39 (br. d, 1H, J=˜8.8,CHCHC≡N), 3.26-3.21 (m, 2H, ArCHCHCH₂Ar, ArCHCHCH₂Ar), 3.03 (dd, 1H,J=18.1, 7.6, CH₂CHCHC≡N), 2.75 (dd, 1H, J=12.7, 2.6, ArCHCH₂NH), 2.62(dd, 1H, J=12.7, 8.5, ArCHCH₂NH), 2.35 (d, 1H, J=18.1, CH₂CHCHC≡N), 2.31(s, 3H, NCH₃), 2.19 (s, 3H, ArCH₃), 2.19 (s, 3H, ArCH₃), 1.83 (dd, 1H,J=15.6, 12.2, ArCHCHCH₂Ar). FTIR (neat film), cm⁻¹ 3303, 2923, 2226,1456, 1108. HRMS (ES) Calcd for C₃₄H₄₂N₅O₆ (MH)⁺: 616.3135, Found:616.3156.

EXAMPLE 6 N-Carbobenzyloxy-L-tryptophan Derivative

Diethylaniline (0.60 μL, 3.8 μmol, 1.1 equiv) was added in one portionto a stirred solution of the amine (1.8 mg, 3.4 μmol, 1 equiv) in THF(0.3 mL) at 0° C. under an argon atmosphere and the solution was stirredfor 5 min. N-Carbobenzyloxy-L-tryptophan (1.5 mg, 4.5 μmol, 1.3 equiv),1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydro-chloride (0.85 mg,4.5 μmol, 1.3 equiv) and 1-hydroxybenzotriazole (0.56 mg, 4.1 μmol, 1.2equiv) were then added separately, each in one portion, to the abovesolution at 0° C. The reaction mixture was warmed to 23° C. over 13 h 55min, then was quenched with saturated aqueous ammonium chloride solution(3 mL). The mixture was diluted with ethyl acetate (10 mL) and thelayers were separated. The aqueous layer was extracted with ethylacetate (10 mL) and the combined organic layer was dried over sodiumsulfate. Concentration in vacuo left a white solid, which was purifiedby flash column chromatography (65% ethyl acetate-hexanes) to give theN-carbobenzyloxy-L-trytophan derivative (2.6 mg, 90%) as a white solid.

¹H NMR (500 MHz, CDCl₃), δ8.20 (br. s, 1H, IndNH), 7.73 (d, 1H, J=7.8,ArH), 7.41 (d, 1H, J=7.8, ArH), 7.35 (br. s, 3H, ArH), 7.23 (m, 2H,ArH), 7.17 (t, 1H, J=7.3, ArH), 7.02 (d, 1H, J=2.4, ArH), 5.71 (br. s,1H, NHCO), 5.58 (s, 1H, ArOH), 5.51 (s, 1H, ArOH), 5.42 (d, 1H, J=7.8,CbzNH), 5.11 (s, 2H, ArCH₂O), 4.36-4.32 (br. m, 1H), 4.04 (d, 1H, J=1.5,CHC≡N), 3.84-3.81 (br. m, 1H), 3.74 (s, 3H, ArOCH₃), 3.70 (s, 6H,2×ArOCH₃), 3.57 (s, 3H, ArOCH₃), 3.24-3.21 (br. m, 2H), 3.16 (dd, 1H,J=15.6, 2.4, ArCHCHCH₂Ar), 3.09-3.04 (m, 3H), 2.83-2.80 (br. m, 1H),2.68-2.61 (br. m, 2H), 2.24 (s, 3H, NCH₃), 2.19 (s, 3H, ArCH₃), 2.17 (s,3H, ArCH₃), 1.97 (d, 1H, J=18.6, CH₂CHCHC≡N, 1.74 (dd, 1H, J=15.6, 12.2,ArCHCHCH₂). FTIR (neat film), cm⁻¹ 3344, 2923, 1708, 1672, 1456. HRMS(ES) Calcd for C₄₇H₅₃N₆O₉ (MH)⁺: 845.3874, Found: 845.391 0.

EXAMPLE 7 Indole-4-carboxylic Acid Amide Derivative

Indole-4-carboxylic acid (0.51 mg, 3.2 μmol, 1.5 equiv), was added inone portion to a stirred solution of the amine (1.1 mg, 2.1 μmol, 1equiv) in dichloromethane (0.15 mL) at 23° C. under an argon atmosphereand the solution was stirred for 10 min.N-Cyclohexylcarbodiimide-N′-propyloxymethyl polystyrene (ArgonautTechnologies, 1.13 mmol/g, 3.7 mg, 4.2 μmol, 2.0 equiv) was then added,in one portion, to the above solution at 23° C. The reaction mixture wasstirred gently at 23° C. for 18 h 20 min, then was purified by flashcolumn chromatography (80% ethyl acetate-hexanes) to give theindole-4-carboxylic acid amide derivative (0.8 mg, 60%) as a whitesolid.

¹H NMR (500 MHz, CDCl₃), δ8.19 (s, 1H, ArNH), 7.40 (d, 1H, J=7.8, ArH),7.16 (t, 1H, J=3.0, ArH), 6.97 (t, 1H, J=7.8, ArH), 6.69 (bt, 1H,J=˜2.0, ArH), 6.52 (d, 1H, J=7.3, ArH), 6.00 (d, 1H, J=6.3, NH), 5.71(s, 1H, ArOH), 5.59 (s, 1H, ArOH), 4.34 (br. s, 1H, CHCH₂NHCO), 4.28(ddd, 1H, J=13.7, 8.3, 2.0, CH₂NHCO), 4.22 (bd, 1H, J=˜2.0, ArCHNCH₃),4.15 (d, 1H, J=2.9, CHC≡N), 3.75 (s, 3H, ArOCH₃),3.74 (s, 3H, ArOCH₃),3.45 (br. d, 1H, J=˜7.3, CHCHC≡N), 3.40 (s, 3H, ArOCH₃), 3.38-3.33 (m,1H (CH₂NHCO), 1H (ArCHCHCH₂Ar)), 3.29 (s, 3H, ArOCH₃), 3.28 (app. dd(obsc.), 1H, J=˜16.1, 2.9, ArCHCHCH₂Ar), 3.08 (dd, 1H, J=18.6, 8.0,CH₂CHCHC≡N), 2.43 (d, 1H, J=18.6, CH₂CHCHC≡N), 2.31 (s, 3H, NCH₃), 2.15(s, 3H, ArCH₃), 2.08 (dd, 1H, J=16.1, 11.7, ArCHCHCH₂Ar), 2.01 (s, 3H,ArCH₃). FTIR (neat film), cm⁻¹ 3393, 2926, 2227, 1641. HRMS (ES) Calcdfor C₃₇H₄₁N₅O₇ (MH)⁺: 668.3084, Found: 668.3062.

EXAMPLE 8 Indole3-carboxylic Acid Amide Derivative

Indole-3-carboxylic acid (0.55 mg, 3.4 μmol, 1.5 equiv), was added inone portion to a stirred solution of the amine (1.2 mg, 2.2 μmol, 1equiv) in dichloromethane (0.2 mL) at 23° C. under an argon atmosphereand the solution was stirred for 10 min.N-Cyclohexylcarbodiimide-N′-propyloxymethyl polystyrene (ArgonautTechnologies, 1.13 mmol/g, 8.0 mg, 9.2 μmol, 4.0 equiv) was then added,in one portion, to the above solution at 23° C. The reaction mixture wasstirred gently at 23° C. for 16 h 45 min, then was purified by flashcolumn chromatography (85% ethyl acetate-hexanes) to give theindole-3-carboxylic acid amide derivative (1.2 mg, 86%) as a whitesolid.

¹H NMR (500 MHz, CDCl₃), δ8.20 (br. s, 1H, ArNH), 7.80 (d, 1H, J=8.3,ArH), 7.32 (app. d, 1H, J=˜7.4, ArH), 7.18 (td, 1H, J=7.4, 1.3, ArH),7.08 (td, 1H, J=7.4, 1.0, ArH), 6.55 (d, 1H, J 2.9, ArH), 5.75 (s, 1H,ArOH), 5.67 (br. d, 1H, J=˜5.9, NH), 5.61 (s, 1H, ArOH), 4.31 (br. s,1H, CHCH₂NHCO), 4.23 (dd, 1H, J=3.0, 1.0, ArCHNCH₃), 4.16-4.12 (m, 2H,CHC≡N, CH₂NHCO), 3.81 (s, 3H, ArOCH₃), 3.74 (s, 3H, ArOCH₃), 3.45 (br.s, 4H, ArOCH₃, CHCHC≡N), 3.37 (ddd, 1H, J=12.0, 2.6, 2.6, ArCHCHCH₂Ar),3.34 (s, 3H, ArOCH₃), 3.30 (ddd, 1H, J=13.7, 3.9, 2.7, CH₂NHCO), 3.26(dd, 1H, J=15.8, 2.6, ArCHCHCH₂Ar), 3.10 (dd, 1H, J=18.5, 7.8,CH₂CHCHC≡N), 2.49 (d, 1H, J=18.5, CH₂CHCHC≡N), 2.34 (s, 3H, NCH₃), 2.14(s, 3H, ArCH₃), 2.06 (s, 3H, ArCH₃), 2.02 (dd, 1H, J=15.8, 11.7,ArCHCHCH₂Ar). FTIR (neat film), cm⁻¹ 3388, 2925, 2225, 1620. HRMS (ES)Calcd for C₃₇H₄₁N₅O₇ (MH)⁺: 668.3084, Found: 668.3062.

EXAMPLE 9 Propionamide Derivative

Diethylaniline (0.60 μL, 4.0 μmol, 1.1 equiv) and propionyl chloride(1.0 mg, 1.0 μL, 10.9 μmol, 3.0 equiv) were added separately, each inone portion, to a stirred solution of the amine (1.9 mg, 3.6 μmol, 1equiv) in dichloromethane (0.2 mL) at 0° C. under an argon atmosphere.The reaction mixture was stirred at 0° C. for 30 min, then was quenchedby the addition of a 1:1 mixture of water and saturated aqueous sodiumhydrogen carbonate solution (4 mL). The mixture was extracted with ethylacetate (2×15 mL) and the combined organic layer was washed with a 1:1mixture of brine solution and saturated aqueous sodium hydrogencarbonate solution (5 mL). The organic layer was dried over sodiumsulfate and concentrated in vacuo to leave a colorless oil. Purificationby flash column chromatography (ethyl acetate→90% ethylacetate-methanol) gave the propionamide derivative (1.9 mg, 90%) as awhite solid. ¹H NMR (500 MHz, CDCl₃), δ5.63 (s, 1H, ArOH), 5.60 (s, 1H,ArOH), 5.21 (d, 1H, J=5.9, NH), 4.20 (d, 1H, J=2.0, ArCHNCH₃), 4.17 (bd,1H, J=2.9, CHCH₂NHCO), 4.03 (d, 1H, J=2.4, CHC≡N, 3.84 (ddd, 1H, J=13.4,8.3, 1.9, CH₂NHCO), 3.78 (s, 3H, ArOCH₃), 3.75 (s, 3H, ArOCH₃), 3.71 (s,3H, ArOCH₃), 3.60 (s, 3H, ArOCH₃), 3.43 (br. d, 1H, J=˜7.8, CHCHC≡N),3.32 (ddd, 1H, J=11.9, 2.7, 2.7, ArCHCHCH₂Ar), 3.24 (dd, 1H, J=15.8,2.7, ArCHCHCH₂Ar), 3.12-3.07 (m, 2H, CH₂CHCHC≡N, CH₂NHCO), 2.50 (d, 1H,J=18.5, CH₂CHCHC≡N), 2.35 (s, 3H, NCH₃), 2.25 (s, 3H, ArCH₃), 2.19 (s,3H, ArCH₃), 1.90 (dd, 1H, J=15.8, 12.0, ArCHCHCH₂Ar), 1.56 (q, 2H,J=7.8, CH₃CH₂CO), 0.74 (t, 3H, J=7.8, CH₃CH₂CO). FTIR (neat film), cm⁻¹3385, 2923, 2226, 1657. HRMS (ES) Calcd for C₃₁H₄₀N₄O₇ (MH)⁺: 581.2975,Found: 581.2948.

EXAMPLE 10 Naphthalene-1-carboxylic Acid Amide Derivative

Diethylaniline (0.27 μL, 1.7 μmol, 1.1 equiv) was added in one portionto a stirred solution of the amine (0.8 mg, 1.5 μmol, 1 equiv) in THF(0.2 mL) at 0° C. under an argon atmosphere and the solution was stirredfor 5 min. 1-Naphthoic acid (0.34 mg, 2.0 μmol, 1.3 equiv),1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (0.38 mg,2.0 μmol, 1.3 equiv) and 1-hydroxybenzotriazole (0.25 mg, 1.8 μmol, 1.2equiv) were then added separately, each in one portion, to the abovesolution at 0° C. The reaction mixture was warmed to 23° C. over 14 h 45min, then was quenched with saturated aqueous ammonium chloride solution(3 mL). The mixture was diluted with ethyl acetate (10 mL) and thelayers were separated. The aqueous layer was extracted with ethylacetate (10 mL) and the combined organic layer was dried over sodiumsulfate. Concentration in vacuo left a white solid, which was purifiedby flash column chromatography (80% ethyl acetate-hexanes) to give thenaphthalene-1-carboxylic acid amide derivative (0.7 mg, 68%) as a whitesolid.

¹H NMR (500 MHz, CDCl₃), δ7.79-7.75 (m, 3H, ArH), 7.45-7.42 (m, 1H,ArH), 7.36 (ddd, 1H, J=7.3, 6.8, 1.2, ArH), 7.19 (dd, 1H, J=7.8, 6.8,ArH), 6.76 (app. d, 1H, J=˜6.8, ArH), 5.71 (s, 1H, ArOH), 5.67 (d, 1H,J=4.9, NH), 5.49 (s, 1H, ArOH), 4.37-4.33 (m, 2H, CH₂NHCO, CHCH₂NHCO),4.17 (d, 1H, J=1.4, ArCHNCH₃), 4.16 (d, 1H, J=2.9, CHC≡N), 3.77 (s, 3H,ArOCH₃), 3.55 (s, 3H, ArOCH₃), 3.45-3.39 (m, 5H, CHCHC≡N, ArOCH₃,CH₂NHCO), 3.34 (ddd, 1H, J=12.0, 2.7, 2.7, ArCHCHCH₂Ar), 3.28 (s, 3H,ArOCH₃), 3.24 (dd, 1H, J=16.1, 2.5, ArCHCHCH₂Ar), 3.05 (dd, 1H, J=18.6,8.0, CH₂CHCHC≡N), 2.43 (d, 1H, J=18.6, CH₂CHCHC≡N), 2.29 (s, 3H, NCH₃),2.20 (s, 3H, ArCH₃), 1.95 (dd, 1H, J=16.1, 11.7, ArCHCHCH₂Ar), 1.88 (s,3H, ArCH₃). FTIR (neat film), cm⁻¹ 3395, 2923, 2226, 1651. HRMS (ES)Calcd for C₃₉H₄₂N₄O₇ (MH)⁺: 679.3132, Found: 679.3106.

EXAMPLE 11 Phenylurea Derivative

Diethylaniline (0.33 μL, 2.1 μmol, 1.1 equiv) and phenyl isocyanate(0.25 μL, 2.3 μmol, 1.2 equiv) were added separately, each in oneportion, to a stirred solution of the amine (1.0 mg, 1.9 μmol, 1 equiv)in dichloromethane (0.2 mL) at 0° C. under an argon atmosphere. Thereaction mixture was stirred at 0° C. for 25 min, then was quenched bythe addition of a 1:1 mixture of water and saturated aqueous sodiumhydrogen carbonate solution (4 mL). The mixture was extracted withdichloromethane (2×10 mL) and the organic layer was dried over sodiumsulfate and concentrated in vacuo to leave a white solid. Purificationby flash column chromatography (70% ethyl acetate) gave the phenylureaderivative (0.9 mg, 74%) as a white solid. ¹H NMR (500 MHz, CDCl₃),δ7.27-7.23 (m, 4H, ArH), 7.01-6.98 (m, 1H, ArH), 6.26 (s, 1H, NH), 5.66(s, 1H, ArOH), 5.53 (s, 1H, ArOH), 4.36 (t, 1H, J=5.8, NH), 4.23-4.21(m, 2H, CHCH₂NHCO, ArCHNCH₃), 4.03 (d, 1H, J=2.4, CHC≡N), 3.81 (s, 6H,2×ArOCH₃), 3.74 (s, 3H, ArOCH₃), 3.59 (s, 3H, ArOCH₃), 3.44 (ddd, 1H,J=13.7, 5.4, 5.4, CH₂NHCO), 3.38 (br. d, 1H, J=7.3, CHCHC≡N), 3.31 (ddd,1H, J=12.7, 2.4, 2.4, ArCHCHCH₂Ar), 3.21 (dd, 1H, J=15.6, 2.9,ArCHCHCH₂Ar), 3.10 (dd, 1H, J=18.5, 7.3, CH₂CHCHC≡N), 2.75-2.69 (m, 2H,CH₂NHCO, CH₂CHCHC≡N), 2.43 (s, 3H, NCH₃), 2.26 (s, 3H, ArCH₃), 2.19 (s,3H, ArCH₃), 1.82 (dd, 1H, J=15.6, 12.5, ArCHCHCH₂Ar). FTIR (neat film),cm⁻¹ 3374, 2923, 2226, 1672. HRMS (ES) Calcd for C₃₅H₄₁N₅O₇ (MH)⁺:644.3084, Found: 644.3110.

EXAMPLE 12 Biphenyl-2-carboxylic Acid Amide Derivative

2-Biphenylcarboxylic acid (0.51 mg, 2.6 μmol, 1.5 equiv) was added inone portion to a stirred solution of the amine (0.9 mg, 1.7 μmol, 1equiv) in dichloromethane (0.2 mL) at 23° C. under an argon atmosphereand the solution was stirred for 10 min.N-Cyclohexylcarbodiimide-N′-propyloxymethyl polystyrene (ArgonautTechnologies, 1.13 mmol/g, 3.0 mg, 3.4 μmol, 2.0 equiv) was then added,in one portion, to the above solution at 23° C. The reaction mixture wasstirred gently at 23° C. for 19 h 30 min, then was purified by flashcolumn chromatography (70% ethyl acetate-hexanes) to give thebiphenyl-2-carboxylic acid amide derivative (0.9 mg, 75%) as a whitesolid.

¹H NMR (500 MHz, CDCl₃), δ7.40 (td, 1H, J=7.3, 1.4, ArH), 7.33-7.29 (m,4H, ArH), 7.24-7.22 (m, 2H, ArH), 7.20 (td, 1H, J=7.3, 1.4, ArH), 6.96(dd, 1H, J=5.8, 1.0, ArH), 5.55 (s, 1H, ArOH), 5.53 (s, 1H, ArOH), 5.52(t, 1H, J=5.8, NH), 4.13 (d, 1H, J=2.5, CHC≡N), 4.03 (t, 1H, J=3.9,CHCH₂NHCO), 3.77 (d, 1H, J=2.5, ArCHNCH₃), 3.73 (s, 3H, ArOCH₃), 3.72(s, 3H, ArOCH₃), 3.58 (s, 3H, ArOCH₃), 3.45 (s, 3H, ArOCH₃), 3.45-3.40(m, 1H, CH₂NHCO), 3.36-3.31 (m, 2H, CH₂NHCO, CHCHC≡N), 3.22-3.18 (m, 2H,ArCHCHCH₂Ar, ArCHCHCH₂Ar), 3.03-2.98 (dd, 1H, J=18.6, 8.3, CH₂CHCHC≡N),2.39 (d, 1H, J=18.6, CH₂CHCHC≡N), 2.27 (s, 3H, NCH₃), 2.20 (s, 3H,ArCH₃), 2.12 (s, 3H, ArCH₃), 1.86 (dd, 1H, J=16.2, 12.2, ArCHCHCH₂Ar).FTIR (neat film), cm⁻¹ 3402, 2930, 2231, 1654. HRMS (ES) Calcd forC₄₁H₄₄N₄O₇ (MH)⁺: 705.3288, Found: 705.3317.

EXAMPLE 13 Phenyl acetyl Derivative

Diethylaniline (0.50 μL, 2.9 μmol, 1.1 equiv) and phenyl acetyl chloride(1.1 μL, 8.0 μmol, 3.0 equiv) were added separately, each in oneportion, to a stirred solution of the amine (1.4 mg, 2.7 μmol, 1 equiv)in dichloromethane (0.15 mL) at 0° C. under an argon atmosphere. Thereaction mixture was stirred at 0° C. for 30 min, then was quenched bythe addition of a 1:1 mixture of water and saturated aqueous sodiumhydrogen carbonate solution (4 mL). The mixture was extracted with ethylacetate (2×15 mL) and the combined organic layer was washed with a 1:1mixture of brine solution and saturated aqueous sodium hydrogencarbonate solution (5 mL). The organic layer was dried over sodiumsulfate and concentrated in vacuo to leave a light yellow oil.Purification by flash column chromatography (80% ethyl acetate-hexanes)gave the phenyl acetyl derivative (1.7 mg, 99%) as a white solid.

¹H NMR (500 MHz, CDCl₃), δ7.21-7.17 (m, 3H, ArH), 6.90-6.89 (m, 2H,ArH), 5.59 (s, 1H, ArOH), 5.41 (t, 1H, J=5.1, NHCO), 5.39 (s, 1H, ArOH),4.17 (d, 1H, J=1.9, ArCHNCH₃), 4.09 (dd, 1H, J=5.4, 2.4, CHCH₂NHCO),4.02 (d, 1H, J=2.5, CHC≡N), 3.75 (s, 3H, ArOCH₃), 3.74 (s, 3H, ArOCH₃),3.71 (s, 3H, ArOCH₃), 3.66 (app. ddd (obsc), 1H, CH₂NHCO), 3.56 (s, 3H,ArOCH₃), 3.39 (br. d, 1H, J=˜8.8, CHCHC≡N), 3.23 (ddd, 1H, J=11.7, 2.4,2.4, ArCHCHCH₂Ar), 3.20-3.15 (m, 2H, CH₂NHCO, ArCHCHCH₂Ar)), 3.15 (d,1H, J=15.1, NHCOCH₂Ar), 3.06 (dd, 1H, J=18.6, 8.3, CH₂CHCHC≡N), 2.96 (d,1H, J=15.1, NHCOCH₂Ar), 2.51 (d, 1H, J=18.6, CH₂CHCHC≡N), 2.32 (s, 3H,NCH₃), 2.24 (s, 3H, ArCH₃), 2.18 (s, 3H, ArCH₃), 1.81 (dd, 1H, J=16.4,12.0, ArCHCHCH₂Ar). FTIR (neat film cm⁻¹ 3385, 2923, 2226, 1667, 1462.HRMS (ES) Calcd for C₃₆H₄₃N₄O₇ (MH)⁺: 643.3132, Found: 643.3130.

EXAMPLE 14 2-Hydroxy-naphthalene-1-carboxylic Acid Amide Derivative

Diethylaniline (0.27 μL, 1.7 μmol, 1.1 equiv) was added in one portionto a stirred solution of the amine (0.8 mg, 1.5 μmol, 1 equiv) in THF(0.2 mL) at 0° C. under an argon atmosphere and the solution was stirredfor 5 min. 2-Hydroxy-1-naphthoic acid (0.37 mg, 2.0 μmol, 1.3 equiv),1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (0.38 mg,2.0 μmol, 1.3 equiv) and 1-hydroxybenzotriazole (0.25 mg, 1.8 μmol, 1.2equiv) were then added separately, each in one portion, to the abovesolution at 0° C. The reaction mixture was warmed to 23° C. over 13 h 20min, then was quenched with saturated aqueous ammonium chloride solution(3 mL). The mixture was diluted with ethyl acetate (10 mL) and thelayers were separated. The aqueous layer was extracted with ethylacetate (10 mL) and the combined organic layer was dried over sodiumsulfate. Concentration in vacuo left a white solid, which was purifiedby flash column chromatography (85% ethyl acetate-hexanes) to give the2-hydroxy-naphthalene-1-carboxylic acid amide derivative (1.1 mg, 100%)as a white solid.

¹H NMR (500 MHz, CDCl₃), δ11.22 (s, 1H, NaphOH), 7.70 (d, 1H, J=8.8,ArH), 7.66 (d, 1H, J=7.8, ArH), 7.28 (d (obsc.), 1H, ArH), 7.23 (t, 1H,J=7.8, ArH), 7.11 (t, 1H, J=7.8, ArH), 7.04 (d, 1H, J=8.8, ArH), 6.10(br. s, 1H, NH), 5.71 (s, 1H, ArOH), 5.46 (s, 1H, ArOH), 4.40 (br. s,1H, CHCH₂NHCO), 4.27 (app. dd, 1H, J=˜13.8, 6.1, CH₂NHCO), 4.17-4.15 (m,2H, ArCHNCH₃, CHC≡N), 3.75 (s, 3H, ArOCH₃), 3.68 (s, 3H, ArOCH₃), 3.54(ddd, 1H, J=14.2, 3.9, 3.9, ArCHCHCH₂Ar), 3.48 (s, 3H, ArOCH₃), 3.42(br. d, 1H, J=˜7.9, CHCHC≡N), 3.38-3.33 (m, 1H, CH₂NHCO), 3.28 (dd, 1H,J=16.2, 2.8, ArCHCHCH₂Ar), 3.15 (s, 3H, ArOCH₃), 2.99 (dd, 1H, J=18.1,8.3, CH₂CHCHC≡N), 2.32 (d, 1H, J=18.1, CH₂CHCHC≡N), 2.25 (s, 3H, NCH₃),2.24 (s, 3H, ArCH₃),2.12 (dd, 1H, J=16.2, 11.8, ArCHCHCH₂Ar), 1.78 (s,3H, ArCH₃). FTIR (neat film), cm⁻¹ 3395, 2923, 2226, 1626. HRMS (ES)Calcd for C₃₉H₄₂N₄O₈ (MH)⁺: 695.3081, Found: 695.3096.

EXAMPLE 15 2-Furoic Acid Amide Derivative

Diethylaniline (0.33 μL, 2.1 μmol, 1.1 equiv) was added in one portionto a stirred solution of the amine (1.0 mg, 1.9 μmol, 1 equiv) in THF(0.15 mL) at 0° C. under an argon atmosphere and the solution wasstirred for 10 min. 2-Furoic acid (0.28 mg, 2.5 μmol, 1.3 equiv),1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (0.48 mg,2.5 μmol, 1.3 equiv) and 1-hydroxybenzotriazole (0.31 mg, 2.3 μmol, 1.2equiv) were then added separately, each in one portion, to the abovesolution at 0° C. The reaction mixture was warmed to 23° C. over 19 h,then was quenched with saturated aqueous ammonium chloride solution (3mL). The mixture was diluted with ethyl acetate (10 mL) and the layerswere separated. The aqueous layer was extracted with ethyl acetate (10mL) and the combined organic layers were dried over sodium sulfate.Concentration in vacuo left a yellow oil, which was purified by flashcolumn chromatography (70% ethyl acetate-hexanes) to give the 2-fuiroicacid amide derivative (1.0 mg, 85%) as a white solid.

¹H NMR (500 MHz, CDCl₃), δ7.18-7.17 (m, 1H, ArH), 6.70 (dd, 1H, J=3.4,1.0, ArH), 6.31 (dd, 1H, J=3.4, 2.0, ArH), 6.07 (d, 1H, J=6.9, NH), 5.63(s, 1H, ArOH), 5.60 (s, 1H, ArOH), 4.28 (br. d, 1H, J=˜2.0, ArCHCH₂NH),4.22 (d, 1H, J=1.5, ArCHNCH₃), 4.10-4.05 (m, 2H, CHC≡N, ArCHCH₂NH), 3.83(s, 3H, ArOCH₃), 3.74 (s, 3H, ArOCH₃), 3.54 (s, 3H, ArOCH₃), 3.45 (br.d, 1H, J=˜7.8, CHCHC≡N), 3.42 (s, 3H, ArOCH₃), 3.32 (ddd, 1H, J=12.0,2.7, 2.7, ArCHCHCH₂Ar), 3.23 (dd, 1H, J=16.1, 2.4, ArCHCHCH₂Ar),3.23-3.19 (1H, m, ArCHCH₂NH), 3.12 (dd, 1H, J=18.6, 8.3, CH₂CHCHC≡N),2.48 (d, 1H, J=18.6, CH₂CHCHC≡N), 2.35 (s, 3H, NCH₃), 2.16 (s, 3H,ArCH₃), 2.15 (s, 3H, ArCH₃), 2.04 (dd, 1H, J=16.1, 11.7, ArCHCHCH₂Ar).FTIR (neat film), cm⁻¹ 3385, 2933, 2226, 1651, 1462. HRMS (ES) Calcd forC₃₃H₃₉N₄O₈ (MH)⁺: 619.2768, Found: 619.2790.

EXAMPLE 16 Indole-3-glyoxamide Derivative

Indole-3-glyoxylic acid (0.55 mg, 2.9 μmol, 1.5 equiv) was added in oneportion to a stirred solution of the amine (1.0 mg, 1.9 μmol, 1 equiv)in dichloromethane (0.15 mL) at 23° C. under an argon atmosphere and thesolution was stirred for 10 min.N-Cyclohexylcarbodiimide-N′-propyloxymethyl polystyrene (ArgonautTechnologies, 1.13 mmol/g, 3.4 mg, 3.8 μmol, 2.0 equiv) was then added,in one portion, to the above solution at 23° C. The reaction mixture wasstirred gently at 23° C. for 15 h 20 min, then was purified by flashcolumn chromatography (70% ethyl acetate-hexanes) to give theindole-3-glyoxamide derivative (0.9 mg, 60%) as a white solid.

¹H NMR (500 MHz, CDCl₃), δ8.85 (d, 1H, J=3.4, ArH), 8.65 (br. s, 1H,ArNH), 8.20 (dd, 1H, J=7.1, 2.1, ArH), 7.42 (dd, 1H, J=6.6, 1.7, ArH),7.32-7.28 (m, 2H, ArH), 7.01 (t, 1H, J=5.7, NH), 5.67 (s, 1H, ArOH),5.55 (s, 1H, ArOH), 4.31 (br. t, 1H, CHCH₂NHCO), 4.19 (d, 1H, J=2.0,ArCHNCH₃), 4.10 (d, 1H, J=2.4, CHC≡N), 3.76 (s, 3H, ArOCH₃), 3.75-3.71(m, 4H, ArOCH₃, CH₂NHCO), 3.61 (s, 3H, ArOCH₃), 3.57 (s, 3H, ArOCH₃),3.45-3.39 (m, 2H, CHCHC≡N, CH₂NHCO), 3.29-3.25 (m, 2H, ArCHCHCH₂Ar,ArCHCHCH₂Ar), 3.09 (dd, 1H, J=18.4, 8.0, CH₂CHCHC≡N), 2.58 (d, 1H,J=18.4, CH₂CHCHC≡N), 2.33 (s, 3H, NCH₃), 2.18 (s, 3H, ArCH₃), 2.10 (dd,1H, J=16.4, 12.0, ArCHCHCH₂Ar), 1.95 (s, 3H, ArCH₃). FTIR (neat film),cm⁻¹ 3370, 2926, 2227, 1675, 1625. HRMS (ES) Calcd for C₃₈H₄₁N₅O₈ (MH)⁺:696.3033, Found: 696.3002.

EXAMPLE 17 Pyrazine-2-carboxylic Acid Amide Derivative

Diethylaniline (0.33 μL, 2.1 μmol, 1.1 equiv) was added in one portionto a stirred solution of the amine (1.0 mg, 1.9 μmol, 1 equiv) in THF(0.2 mL) at 0° C. under an argon atmosphere and the solution was stirredfor 5 min. 2-Pyrazinecarboxylic acid (0.31 mg, 2.5 μmol, 1.3 equiv),1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (0.48 mg,2.5 μmol, 1.3 equiv) and 1-hydroxybenzotriazole (0.31 mg, 2.3 μmol, 1.2equiv) were then added separately, each in one portion, to the abovesolution at 0° C. The reaction mixture was warmed to 23° C. over 17 h 40min, then was quenched with saturated aqueous ammonium chloride solution(3 mL). The mixture was diluted with ethyl acetate (10 mL) and thelayers were separated. The aqueous layer was extracted with ethylacetate (10 mL) and the combined organic layer was dried over sodiumsulfate. Concentration in vacuo left a white solid, which was purifiedby flash column chromatography (80% ethyl acetate-hexanes) to give thepyrazine-2-carboxylic acid amide derivative (1.1 mg, 92%) as a whitesolid.

¹H NMR (500 MHz, CDCl₃), δ9.10 (d, 1H, J=1.5, ArH), 8.60 (d, 1H, J=2.4,ArH), 8.17 (app. t, 1H, J=˜2.0, ArH), 7.33 (t, 1H, J=5.4, NH), 5.65 (s,1H, ArOH), 5.52 (s, 1H, ArOH), 4.33 (br. s, 1H, CHCH₂NHCO), 4.18 (br. s,1H, ArCHNCH₃), 4.11 (d, 1H, J=2.4, CHC≡N), 3.93 (ddd, 1H, J=13.4, 6.7,2.0, CH₂NHCO), 3.75 (s, 3H, ArOCH₃), 3.74 (s, 3H, ArOCH₃), 3.57 (s, 3H,ArOCH₃), 3.49 (ddd, 1H, J=13.4, 4.4, 4.4, CH₂NHCO), 3.45 (obsc. d, 1H,CHCHC≡N), 3.44 (s, 3H, ArOCH₃), 3.29 (br. d, 1H, J=11.7, ArCHCHCH₂Ar),3.20 (dd, 1H, J=16.1, 2.4, ArCHCHCH₂Ar), 3.07 (dd, 1H, J=18.5, 8.3,CH₂CHCHC≡N), 2.55 (d, 1H, J=18.5, CH₂CHCHC≡N), 2.32 (s, 3H, NCH₃), 2.16(s, 3H, ArCH₃), 2.08 (dd, 1H, J=16.1, 12.2, ArCHCHCH₂Ar), 2.08 (s, 3H,ArCH₃). FTIR (neat film), cm⁻¹ 3372, 2922, 2231, 1728, 1676. HRMS (ES)Calcd for C₃₃H₃₈N₆O₇ (MH)⁺: 631.2880, Found: 631.2853.

EXAMPLE 18 Pyruvamide Derivative

Diethylaniline (0.37 μL, 2.3 μmol, 1.1 equiv) was added in one portionto a stirred solution of the amine (1.1 mg, 2.1 μmol, 1 equiv) in THF(0.15 mL) at 0° C. under an argon atmosphere and the solution wasstirred for 5 min. Pyruvic acid (0.18 μL, 2.7 μmol, 1.3 equiv),1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (0.52 mg,2.7 μmol, 1.3 equiv) and 1-hydroxybenzotriazole (0.34 mg, 2.5 μmol, 1.2equiv) were then added separately, each in one portion, to the abovesolution at 0° C. The reaction mixture was warmed to 23° C. over 14 h 15min, then was quenched with saturated aqueous ammonium chloride solution(3 mL). The mixture was diluted with ethyl acetate (10 mL) and thelayers were separated. The aqueous layer was extracted with ethylacetate (10 mL) and the combined organic layer was dried over sodiumsulfate. Concentration in vacuo left a white solid, which was purifiedby flash column chromatography (70% ethyl acetate-hexanes) to give thepyruvamide derivative (1.2 mg, 96%) as a white solid.

¹H NMR (500 MHz, CDCl₃), δ6.45 (t, 1H, J=5.6, NHCO), 5.62 (s, 1H, ArOH),5.56 (s, 1H, ArOH), 4.24 (dd, 1H, J=4.1, 1.9, CHCH₂NHCO), 4.17 (dd, 1H,J=3.1, 1.2, ArCHNCH₃), 4.03 (d, 1H, J=2.9, CHC≡N), 3.81 (s, 3H, ArOCH₃),3.76 (s, 3H, ArOCH₃), 3.67 (s, 3H, ArOCH₃), 3.66 (app. ddd (obsc), 1H,CH₂NHCO), 3.60 (s, 3H, ArOCH₃), 3.42 (br. d, 1H, J=˜8.8, CHCHC≡N), 3.31(ddd, 1H, J=13.6, 5.6, 4.1, CH₂NHCO), 3.26-3.21 (m, 2H, ArCHCHCH₂Ar,ArCHCHCH₂Ar), 3.07 (dd, 1H, J=18.6, 8.3, CH₂CHCHC≡N), 2.49 (d, 1H,J=18.6, CH₂CHCHC≡N), 2.32 (s, 3H, NCH₃), 2.24 (s, 3H, COCH₃), 2.20 (s,3H, ArCH₃), 2.17 (s, 3H, ArCH₃), 1.93 (dd, 1H, J=16.4, 12.0,ArCHCHCH₂Ar). FTIR (neat film), cm⁻¹ 3378, 2937, 2251, 1720, 1682, 1463.HRMS (ES) Calcd for C₃₁H₃₉N₄O₉ (MH)⁺: 595.2768, Found: 595.2787.

EXAMPLE 19 Benzamide Derivative

Diethylaniline (0.33 μL, 2.1 μmol, 1.1 equiv) and benzoyl chloride (0.66μL, 5.7 μmol, 3.0 equiv) were added separately, each in one portion, toa stirred solution of the amine (1.0 mg, 1.9 μmol, 1 equiv) indichloromethane (0.15 mL) at 0° C. under an argon atmosphere. Thereaction mixture was stirred at 0° C. for 30 min, then was quenched bythe addition of a 1:1 mixture of water and saturated aqueous sodiumhydrogen carbonate solution (4 mL). The mixture was extracted with ethylacetate (2×15 mL) and the combined organic layer was washed with a 1:1mixture of brine solution and saturated aqueous sodium hydrogencarbonate solution (4 mL). The organic layer was dried over sodiumsulfate and concentrated in vacuo to leave a white solid. Purificationby flash column chromatography (70% ethyl acetate-hexanes) gave thebenzamide derivative (1.1 mg, 92%) as a white solid.

¹H NMR (500 MHz, CDCl₃), δ7.39-7.36 (m, 1H, ArH), 7.23-7.20 (m, 2H,ArH), 7.06-7.04 (m, 2H, ArH), 5.99 (br. d, 1H, J=˜6.3, NHCO), 5.67 (s,1H, ArOH), 5.65 (s, 1H, ArOH), 4.31 (br. d, 1H, J=˜2.4, CHCH₂NHCO), 4.24(dd, 1H, J=2.9, 1.5, ArCHNCH₃), 4.20 (ddd, 1H, J=13.7, 8.3, 2.0,CH₂NHCO), 4.10 (d, 1H, J=2.9, CHC≡N), 3.82 (s, 3H, ArOCH₃), 3.74 (s, 3H,ArOCH₃), 3.45 (br. d, 1H, J=˜8.8, CHCHC≡N), 3.44 (s, 3H, ArOCH₃), 3.38(s, 3H, ArOCH₃), 3.36 (app. ddd (obsc), 1H, ArCHCHCH₂Ar), 3.32-3.27 (m,2H, CH₂NHCO, ArCHCHCH₂Ar), 3.11 (dd, 1H, J=18.5, 7.8, CH₂CHCHC≡N), 2.43(d, 1H, J=18.5, CH₂CHCHC≡N), 2.33 (s, 3H, NCH₃), 2.16 (s, 3H, ArCH₃),2.07 (s, 3H, ArCH₃), 2.03 (dd, 1H, J=15.9, 12.0, ArCHCHCH₂Ar). FTIR(neat film), cm⁻¹ 3395, 2933, 2226, 1656, 1462, 1056. HRMS (ES) Calcdfor C₃₅H₄₁N₄O₇ (MH)⁺: 629.2975, Found: 629.3002.

EXAMPLE 20 Carbamic acid benzyl ester Derivative

Diethylaniline (0.27 μL, 1.7 μmol, 1.1 equiv) and benzyl chloroformate(0.65 μL, 4.6 μmol, 3.0 equiv) were added separately, each in oneportion, to a stirred solution of the amine (0.8 mg, 1.5 μmol, 1 equiv)in dichloromethane (0.1 mL) at 0° C. under an argon atmosphere. Thereaction mixture was stirred at 0° C. for 30 min, then was quenched bythe addition of a 1:1 mixture of water and saturated aqueous sodiumhydrogen carbonate solution (3 mL). The mixture was extracted with ethylacetate (2×10 mL) and the combined organic layer was washed with a 1:1mixture of brine solution and saturated aqueous sodium hydrogencarbonate solution (5 mL). The organic layer was dried over sodiumsulfate and concentrated in vacuo to leave a light yellow oil.Purification by flash column chromatography (60% ethyl acetate-hexanes)gave the carbamic acid benzyl ester derivative (1.0 mg, 99%) as a whitesolid.

¹H NMR (500 MHz, CDCl₃), δ7.30-7.27 (m, 3H, ArH), 7.14-7.12 (m, 2H,ArH), 5.58 (s, 1H, ArOH), 5.52 (s, 1H, ArOH), 4.80 (s, 2H, OCH₂Ar), 4.42(br. d, 1H, J=˜5.4, NHCO), 4.17-4.15 (m, 2H, ArCHNCH₃, CHCH₂NHCO), 3.99(d, 1H, J 2.0, CHC≡N), 3.74 (s, 3H, ArOCH₃), 3.64 (s, 3H, ArOCH₃), 3.60(s, 3H, ArOCH₃), 3.58 (app. ddd (obsc), 1H, CH₂NHCO), 3.58 (s, 3H,ArOCH₃), 3.38 (br. d, 1H, J=˜7.3, CHCHC≡N), 3.28 (ddd, 1H, J=12.0, 2.7,2.7 ArCHCHCH₂Ar), 3.20 (dd, 1H, J=15.8, 2.4, ArCHCHCH₂Ar), 3.14 (ddd,1H, J=13.4, 3.9, 3.9, CH₂NHCO), 3.07 (dd, 1H, J=18.6, 7.8, CH₂CHCHC≡N),2.47 (d, 1H, J=18.6, CH₂CHCHC≡N), 2.33 (s, 3H, NCH₃), 2.20 (s, 3H,ArCH₃), 2.10 (s, 3H, ArCH₃), 1.90 (dd, 1H, J=15.8, 12.0, ArCHCHCH₂Ar).FTIR (neat film), cm⁻¹ 3405, 2923, 2226, 1713, 1462. HRMS (ES) Calcd forC₃₆H₄₃N₄O₈ (MH)⁺: 659.3081, Found: 659.3110.

EXAMPLE 21 Carbamic acid 4-methoxy-phenyl ester Derivative

Diethylaniline (0.33 μL, 2.1 μmol, 1.1 equiv) and 4-methoxyphenylchloroformate (0.85 μL, 5.7 μmol, 3.0 equiv) were added separately, eachin one portion, to a stirred solution of the amine (1.0 mg, 1.9 μmol, 1equiv) in dichloromethane (0.15 mL) at 0° C. under an argon atmosphere.The reaction mixture was stirred at 0° C. for 40 min, then was quenchedby the addition of a 1:1 mixture of water and saturated aqueous sodiumhydrogen carbonate solution (3 mL). The mixture was extracted with ethylacetate (2×10 mL) and the combined organic layer was washed with a 1:1mixture of brine solution and saturated aqueous sodium hydrogencarbonate solution (5 mL). The organic layer was dried over sodiumsulfate and concentrated in vacuo to leave a colorless oil. Purificationby flash column chromatography (50%→70% ethyl acetate-hexanes) gave thecarbamic acid 4-methoxyphenyl ester derivative (0.9 mg, 70%) as a whitesolid.

¹H NMR (500 MHz, CDCl₃), δ6.75 (d, 1H, J=9.3, ArH), 6.67 (d, 1H, J=8.8,ArH), 5.61 (s, 1H, ArOH), 5.57 (s, 1H, ArOH), 4.67 (app. d, 1H, J=6.8,NHCO), 4.21 (d, 1H, J=ArCHNCH₃), 4.19 (br. d, 1H, J=˜2.5, CHCH₂NHCO),4.05 (d, 1H, J=2.4, CHC≡N), 3.75 (s, 3H, ArOCH₃), 3.74 (s, 3H, ArOCH₃),3.71 (s, 3H, ArOCH₃), 3.67 (app. ddd (obsc), 1H, CH₂NHCO), 3.63 (s, 3H,ArOCH₃), 3.62 (s, 3H, ArOCH₃), 3.43 (br. d, 1H, J=˜7.8, CHCHC≡N), 3.34(ddd, 1H, J=12.0, 2.5, 2.5, ArCHCHCH₂Ar), 3.26 (dd, 1H, J=15.8, 2.8,ArCHCHCH₂Ar), 3.16-3.10 (m, 2H, CH₂NHCO, CH₂CHCHC≡N), 2.51 (d, 1H,J=18.6, CH₂CHCHC≡N), 2.37 (s, 3H, NCH₃), 2.22 (s, 3H, ArCH₃), 2.17 (s,3H, ArCH₃), 2.01 (dd, 1H, J=15.8, 12.0, ArCHCHCH₂Ar). FTIR (neat film),cm⁻¹ 3508, 3385, 2923, 2226, 1733, 1195. HRMS (ES) Calcd for C₃₆H₄₃N₄O₉(MH)⁺: 675.3030, Found: 675.3019.

EXAMPLE 22 Pyridine-2-carboxylic Acid Amide Derivative

Diethylaniline (0.27 μL, 1.7 μmol, 1.1 equiv) was added in one portionto a stirred solution of the amine (0.8 mg, 1.5 μmol, 1 equiv) in THF(0.15 mL) at 0° C. under an argon atmosphere and the solution wasstirred for 10 min. Picolinic acid (0.24 mg, 2.0 μmol, 1.3 equiv),1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (0.38 mg,2.0 μmol, 1.3 equiv) and 1-hydroxybenzotriazole (0.25 mg, 1.8 μmol, 1.2equiv) were then added separately, each in one portion, to the abovesolution at 0° C. The reaction mixture was warmed to 23° C. over 17 h 45min, then was quenched with saturated aqueous ammonium chloride solution(3 mL). The mixture was diluted with ethyl acetate (10 mL) and thelayers were separated. The aqueous layer was extracted with ethylacetate (10 mL) and the combined organic layer was dried over sodiumsulfate. Concentration in vacuo left a yellow oil, which was purified byflash column chromatography (70% ethyl acetate-hexanes) to give thepyridine-2-carboxylic acid amide derivative (1.0 mg, 100%) as a whitesolid.

¹H NMR (500 MHz, CDCl₃), δ8.20-8.19 (m, 1H, ArH), 7.92 (dd, 1H, J=7.8,1.0, ArH), 7.71 (ddd, 1H, J=7.6, 7.6, 1.5, ArH), 7.68 (t, 1H, J=5.6,NH), 7.30 (m, 1H, ArH), 5.65 (s, 1H, ArOH), 5.52 (s, 1H, ArOH), 4.28 (t,1H, J=3.2, ArCHCH₂NH), 4.22 (dd, 1H, J=3.0, 1.0, ArCHNCH₃), 4.13 (d, 1H,J=2.5, CHC≡N), 3.85 (ddd, 1H, J=13.2, 6.6, 3.0, ArCHCH₂NH), 3.75 (s, 3H,ArOCH₃), 3.73 (s, 3H, ArOCH₃), 3.55 (s, 3H, ArOCH₃), 3.51-3.45 (m, 1H,ArCHCH₂NH), 3.44 (s, 3H, ArOCH₃), 3.44 (br. d, 1H, J=˜7.8, CHCHC≡N),3.28 (ddd, 1H, J=12.0, 3.0, 3.0, ArCHCHCH₂Ar), 3.22 (dd, 1H, J=15.7,2.4, ArCHCHCH₂Ar), 3.08 (dd, 1H, J=18.6, 8.1, CH₂CHCHCH₂Ar), 2.58 (d,1H, J=18.6, CH₂CHCHC≡N), 2.31 (s, 3H, NCH₃), 2.16 (s, 3H, ArCH₃), 2.16(dd, 1H (obsc), ArCHCHCH₂Ar), 2.09 (s, 3H, ArCH₃). HRMS (ES) Calcd forC₃₄H₄₀N₅O₇ (MH)⁺: 630.2927, Found: 630.2904.

EXAMPLE 23 Indole-2-carboxylic Acid Amide Derivative

Diethylaniline (0.33 μL, 2.1 μmol, 1.1 equiv) was added in one portionto a stirred solution of the amine (1.0 mg, 1.9 μmol, 1 equiv) in THF(0.15 mL) at 0° C. under an argon atmosphere and the solution wasstirred for 5 min. Indole-2-carboxylic acid (0.40 mg, 2.5 μmol, 1.3equiv), 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride(0.48 mg, 2.5 μmol, 1.3 equiv) and 1-hydroxybenzothiazole (0.31 mg, 2.3μmol, 1.2 equiv) were then added separately, each in one portion, to theabove solution at 0° C. The reaction mixture was warmed to 23° C. over15 h 30 min, then was quenched with saturated aqueous ammonium chloridesolution (3 mL). The mixture was diluted with ethyl acetate (10 mL) andthe layers were separated. The aqueous layer was extracted with ethylacetate (10 mL) and the combined organic layer was dried over sodiumsulfate. Concentration in vacuo left a white solid, which was purifiedby flash column chromatography (60% ethyl acetate-hexanes) to give theindole-2-carboxylic acid amide derivative (1.1 mg, 87%) as a whitesolid.

¹H NMR (500 MHz, CDCl₃), δ8.80 (s, 1H, ArNH), 7.65 (d, 1H, J=8.7, ArH),7.31 (d, 1H, J=8.3, ArH), 7.23 (ddd, 1H, J=6.9, 6.9, 1.0, ArH), 7.12(ddd, 1H, J=6.9, 6.9, 1.0, ArH), 5.89 (dd, 1H, J=8.3, 2.5, NH), 5.69 (s,1H, ArOH), 5.68 (s, 1H, ArOH), 5.58 (d, 1H, J=1.5, ArH), 4.31 (br. s,1H, ArCHCH₂NH), 4.26 (dd, 1H, J=3.0, 1.0, ArCHNCH₃), 4.16 (ddd, 1H,J=13.6, 8.5, 2.4, ArCHCH₂NH), 4.10 (d, 1H, J=2.5, CHC≡N), 3.80 (s, 3H,ArOCH₃), 3.74 (s, 3H, ArOCH₃), 3.56 (s, 3H, ArOCH₃), 3.48 (br. d, 1H,J=˜8.5, CHCHC≡N), 3.37-3.33 (m, 2H, ArCHCH₂NH, ArCHCHCH₂Ar), 3.32 (s,3H, ArOCH₃), 3.25 (dd, 1H, J=16.1, 2.9, ArCHCHCH₂Ar), 3.14 (dd, 1H,J=18.5, 8.3, CH₂CHCHC≡N), 2.55 (d, 1H, J=18.5, CH₂CHCHC≡N), 2.35 (s,3H,NCH₃), 2.15 (s, 3H, ArCH₃), 2.13 (s, 3H, ArCH₃), 2.01 (dd, 1H, J=16.1,12.2, ArCHCHCH₂Ar). FTIR (neat film), cm⁻¹ 3405, 3292, 2923, 2226, 1646,1544, 1056. HRMS (ES) Calcd for C₃₇H₄₂N₅O₇ (MH)⁺: 668.3084, Found:668.3112.

EXAMPLE 24 Isoquinoline-1-carboxylic Acid Amide Derivative

1-Isoquinolinecarboxylic acid (0.59 mg, 3.4 μmol, 1.5 equiv) was addedin one portion to a stirred solution of the amine (1.2 mg, 2.3 μmol, 1equiv) in dichloromethane (0.15 mL) at 23° C. under an argon atmosphereand the solution was stirred for 10 min.N-Cyclohexylcarbodiimide-N′-propyloxymethyl polystyrene (ArgonautTechnologies, 1.13 mmol/g, 4.0 mg, 4.6 μmol, 2.0 equiv) was then added,in one portion, to the above solution at 23° C. The reaction mixture wasstirred gently at 23° C. for 21 h, then was purified by flash columnchromatography (85% ethyl acetate-hexanes) to give theisoquinoline-1-carboxylic acid amide derivative (1.1 mg, 71%) as a whitesolid.

¹H NMR (500 MHz, CDCl₃), δ9.30 (d, 1H, J=8.8, ArH), 8.09 (d, 1H, J=5.4,ArH), 7.77 (d, 1H, J=8.3, ArH), 7.76 (t, 1H, J=6.4, NH), 7.68-7.65 (m,2H, ArH), 7.60-7.56 (m, 1H, ArH), 5.71 (s, 1H, ArOH), 5.55 (s, 1H,ArOH), 4.37 (t, 1H, J=3.3, CHCH₂NHCO), 4.20 (app. d, 1H, J=˜2.0,ArCHNCH₃), 4.16 (d, 1H, J=2.5, CHC≡N), 3.92 (ddd, 1H, J=13.6, 7.1, 3.2,CH₂NHCO), 3.75 (s, 3H, ArOCH₃), 3.71 (s, 3H, ArOCH₃), 3.51 (ddd, 1H,J=13.6, 4.4, 4.4, CH₂NHCO), 3.44 (s, 4H, ArOCH₃, CHCHC≡N), 3.40 (s, 3H,ArOCH₃), 3.31 (ddd, 1H, J=11.8, 2.7, 2.7, ArCHCHCH₂Ar), 3.24 (dd, 1H,J=15.6, 2.4, ArCHCHCH₂Ar), 3.07 (dd, 1H, J=18.4, 7.6, CH₂CHCHC≡N), 2.61(d, 1H, J=18.4, CH₂CHCHC≡N), 2.31 (s, 3H, NCH₃), 2.28 (dd, 1H, J=15.6,11.7, ArCHCHCH₂Ar), 2.14 (s, 3H, ArCH₃), 1.89 (s, 3H, ArCH₃). FTIR (neatfilm), cm⁻¹ 3376, 2926, 2230, 1665. HRMS (ES) Calcd for C₃₈H₄₁N₅O₇(MH)⁺: 680.3084, Found: 680.3112.

EXAMPLE 25 5-Fluoro-indole-2-carboxylic Acid Amide Derivative

5-Fluoro-2-indolecarboxylic acid (0.51 mg, 2.9 μmol, 1.5 equiv) wasadded in one portion to a stirred solution of the amine (1.0 mg, 1.9μmol, 1 equiv) in dichloromethane (0.2 mL) at 23° C. under an argonatmosphere and the solution was stirred for 10 min.N-Cyclohexylcarbodiimide-N′-propyloxymethyl polystyrene (ArgonautTechnologies, 1.13 mmol/g, 3.4 mg, 3.8 μmol, 2.0 equiv) was then added,in one portion, to the above solution at 23° C. The reaction mixture wasstirred gently at 23° C. for 16 h 40 min, then was purified by flashcolumn chromatography (80% ethyl acetate-hexanes) to give the5-fluoro-indole-2-carboxylic acid amide derivative (1.1 mg, 85%) as awhite solid.

¹H NMR (500 MHz, CDCl₃), δ8.85 (br. s, 1H, ArNH), 7.32 (dd, 1H, J=9.2,2.4, ArH), 7.22 (dd, 1H, J=10.8, 4.4, ArH), 6.98 (td, 1H, J=8.8, 2.4,ArH), 5.87 (br. d, 1H, J=˜6.9, NH), 5.72 (s, 1H, ArOH), 5.69 (s, 1H,ArOH), 5.43 (br. d, 1H, J=˜2.0, ArH), 4.30 (br. s, 1H, CHCH₂NHCO), 4.25(br. d, 1H, J=˜2.9, ArCHNCH₃), 4.20 (ddd, 1H, J=13.4, 8.8, 2.0,CH₂NHCO), 4.09 (d, 1H, J=2.5, CHC≡N), 3.84 (s, 3H, ArOCH₃), 3.73 (s, 3H,ArOCH₃), 3.55 (s, 3H, ArOCH₃), 3.47 (br. d, 1H, J=˜8.8, CHCHC≡N), 3.35(ddd, 1H, J=11.7, 2.7, 2.7, ArCHCHCH₂Ar), 3.28 (obsc. ddd, 1H, CH₂NHCO),3.27 (s, 3H, ArOCH₃), 3.24 (dd, 1H, J=16.1, 2.5, ArCHCHCH₂Ar), 3.13 (dd,1H, J=18.6, 8.3, CH₂CHCHC≡N), 2.53 (d, 1H, J=18.6, CH₂CHCHC≡N), 2.35 (s,3H, NCH₃), 2.15 (s, 3H, ArCH₃), 2.11 (s, 3H, ArCH₃), 1.98 (dd, 1H,J=16.1, 12.0, ArCHCHCH₂Ar). FTIR (neat film), cm⁻¹ 3398, 2925, 2225,1645. HRMS (ES) Calcd for C₃₇H₄₀N₅O₇F (MH)⁺: 686.2990, Found: 686.3020.

EXAMPLE 26 Phenylpyruvamide Derivative

Diethylaniline (0.43 μL, 2.7 μmol, 1.1 equiv) was added in one portionto a stirred solution of the amine (1.3 mg, 2.5 μmol, 1 equiv) in THF(0.25 mL) at 0° C. under an argon atmosphere and the solution wasstirred for 5 min. Phenylpyruvic acid (0.53 mg, 3.2 μmol, 1.3 equiv),1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (0.62 mg,3.2 μmol, 1.3 equiv) and 1-hydroxybenzotriazole (0.40 mg, 3.0 μmol, 1.2equiv) were then added separately, each in one portion, to the abovesolution at 0° C. The reaction mixture was warmed to 23° C. over 13 h 40min, then was quenched with saturated aqueous ammonium chloride solution(3 mL). The mixture was diluted with ethyl acetate (10 mL) and thelayers were separated. The aqueous layer was extracted with ethylacetate (10 mL) and the combined organic layer was dried over sodiumsulfate. Concentration in vacuo left a white solid, which was purifiedby flash column chromatography (70% ethyl acetate-hexanes) to give thephenylpyruvamide derivative (1.4 mg, 84%) as a white solid.

¹H NMR (500 MHz, CDCl₃), δ7.27-7.23 (m, 3H, ArH), 7.04-7.02 (m, 2H,ArH), 6.56 (br. d, 1H, J=˜4.9, NH), 5.61 (s, 1H, ArOH), 5.54 (s, 1H,ArOH), 4.25 (br. d, 1H, J=˜2.0, CHCH₂NHCO), 4.17 (br. d, 1H, J=˜2.4,ArCHNCH₃), 4.00 (d, 1H, J=2.4, CHC≡N), 3.90 (AB system, 2H,ArCH₂COCONH), 3.84 (obsc. ddd, 1H, J=13.7, 8.5, 1.9, CH₂NHCO), 3.74 (s,3H, ArOCH₃), 3.60 (s, 3H, ArOCH₃), 3.54 (s, 3H, ArOCH₃), 3.52 (s, 3H,ArOCH₃), 3.41 (br. d, 1H, J=˜7.8, CHCHC≡N), 3.27 (ddd, 1H, J=11.7, 2.5,2.5, ArCHCHCH₂Ar), 3.21 (dd, 1H, J=16.1, 2.4, ArCHCHCH₂Ar), 3.17 (ddd,1H, J=13.7, 3.9, 3.9, CH₂NHCO), 3.07 (dd, 1H, J=18.6, 8.3, CH₂CHCHC≡N),2.42 (d, 1H, J=18.6, CH₂CHCHC≡N), 2.32 (s, 3H, NCH₃), 2.18 (s, 3H,ArCH₃), 2.11 (s, 3H, ArCH₃), 1.94 (dd, 1H, J=16.1, 12.0, ArCHCHCH₂Ar).FTIR (neat film), cm⁻¹ 3367,2929, 2229, 1723, 1681. HRMS (ES) Calcd forC₃₇H₄₂N₄O₈ (MH)⁺: 671.3081, Found: 671.3112.

EXAMPLE 27 2-Fluorobenzamide Derivative

2-Fluorobenzoic acid (0.36 mg, 2.6 μmol, 1.5 equiv) was added in oneportion to a stirred solution of the amine (0.9 mg, 1.7 μmol, 1 equiv)in dichloromethane (0.2 mL) at 23° C. under an argon atmosphere and thesolution was stirred for 10 min.N-Cyclohexylcarbodiimide-N′-propyloxymethyl polystyrene (ArgonautTechnologies, 1.13 mmol/g, 3.0 mg, 3.4 μmol, 2.0 equiv) was then added,in one portion, to the above solution at 23° C. The reaction mixture wasstirred gently at 23° C. for 13 h, then was purified by flash columnchromatography (70% ethyl acetate-hexanes) to give the 2-fluorobenzamidederivative (1.1 mg, 99%) as a white solid.

¹H NMR (500 MHz, CDCl₃), δ7.78 (td, 1H, J=7.8, 1.9, ArH), 7.38-7.34 (m,1H, ArH), 7.13 (td, 1H, J=7.8, 1.0, ArH), 6.85-6.81 (m, 1H, ArH), 6.35(br. s, 1H, NH), 5.68 (s, 1H, ArOH), 5.56 (s, 1H, ArOH), 4.32 (br. d,1H, J=˜1.9, CHCH₂NHCO), 4.20 (dd, 1H, J=12.9, 1.0, ArCHNCH₃), 4.12 (d,1H, J=2.5, CHC≡N), 4.09-4.05 (m, 1H, CH₂NHCO), 3.75 (s, 6H, 2×ArOCH₃),3.59 (s, 3H, ArOCH₃), 3.46 (obsc. ddd, 1H, J=14.2, 3.7, 3.7, CH₂NHCO),3.44 (br. d, 1H, J=˜8.8, CHCHC≡N), 3.37 (s, 3H, ArOCH₃), 3.30 (br. d,1H, J=˜11.7, ArCHCHCH₂Ar), 3.26 (dd, 1H, J=15.6, 2.7, ArCHCHCH₂Ar), 3.06(dd, 1H, J=18.5, 8.0, CH₂CHCHC≡N), 2.43 (d, 1H, J=18.5, CH₂CHCHC≡N),2.30 (s, 3H, NCH₃), 2.19 (s, 3H, ArCH₃), 2.11 (dd, 1H, J=15.6, 11.7,ArCHCHCH₂Ar), 2.00 (s, 3H, ArCH₃). FTIR (neat film), cm⁻¹ 3403, 2932,2231, 1649. HRMS (ES) Calcd for C₃₅H₃₉N₄O₇F (MH)⁺: 647.2881, Found:647.2903.

EXAMPLE 28 Quinoline-2-carboxylic Acid Amide Derivative

Quinaldic acid (0.50 mg, 2.9 μmol, 1.5 equiv) was added in one portionto a stirred solution of the amine (1.0 mg, 1.9 μmol, 1 equiv) indichloromethane (0.2 mL) at 23° C. under an argon atmosphere and thesolution was stirred for 10 min.N-Cyclohexylcarbodiimide-N′-propyloxymethyl polystyrene (ArgonautTechnologies, 1.13 mmol/g, 3.4 mg, 3.8 μmol, 2.0 equiv) was then added,in one portion, to the above solution at 23° C. The reaction mixture wasstirred gently at 23° C. for 16 h 50 min, then was purified by flashcolumn chromatography (70% ethyl acetate-hexanes) to give thequinoline-2-carboxylic acid amide derivative (1.1 mg, 85%) as a whitesolid.

¹H NMR (500 MHz, CDCl₃), δ8.23 (d, 1H, J=8.3, ArH), 8.13 (d, 1H, J=8.3,ArH), 7.95 (t, 1H, J=5.6, NH), 7.84 (d, 1H, J=8.1, ArH), 7.81 (d, 1H,J=8.1, ArH), 7.72 (ddd, 1H, J=8.3, 6.8, 1.4, ArH), 7.59 (ddd, 1H, J=8.3,6.8, 1.0, ArH), 5.71 (br. s, 1H, ArOH), 5.44 (s, 1H, ArOH), 4.39 (t, 1H,J=3.9, CHCH₂NHCO), 4.19 (d, 1H, J=2.4, CHC≡N), 4.17 (d, 1H, J=1.5,ArCHNCH₃), 3.81-3.77 (m, 4H, CH₂NHCO, ArOCH₃), 3.61-3.57 (s, 7H,CH₂NHCO, 2×ArOCH₃), 3.42 (br. d, 1H, CHCHC≡N), 3.38 (s, 3H, ArOCH₃),3.31-3.26 (m, 2H, ArCHCHCH₂Ar, CH₂CHCHC≡N), 3.08 (dd, 1H, J=18.6, 8.3,CH₂CHCHC≡N), 2.74 (d, 1H, J=18.6, CH₂CHCHC≡N), 2.27 (s, 3H, NCH₃), 2.21(s, 3H, ArCH₃), 2.16 (dd, 1H, J=16.1, 12.2, ArCHCHCH₂Ar), 1.94 (s, 3H,ArCH₃). FTIR (neat film), cm⁻¹ 3385, 2933, 2226, 1667. HRMS (ES) Calcdfor C₃₈H₄₁N₅O₇ (MH)⁺: 680.3084, Found: 680.3094.

EXAMPLE 29 4-Methylphenyl sulfonamide Derivative

Diethylaniline (0.33 μL, 2.1 μmol, 1.1 equiv) was added in one portionto a stirred solution of the amine (1.0 mg, 1.9 μmol, 1 equiv) in THF(0.1 mL) at 0° C. under an argon atmosphere and the solution was stirredfor 5 min. p-Toluenesulfonyl chloride (0.73 mg, 3.8 μmol, 2.0 equiv) wasthen added in one portion to the above solution at 0° C. The reactionmixture was stirred at 0° C. for 1 hr and then quenched with a 1:1mixture of saturated aqueous sodium hydrogen carbonate solution andwater (4 mL). The mixture was diluted with ethyl acetate (10 mL) and thelayers were separated. The aqueous layer was extracted with ethylacetate (10 mL) and the combined organic layer was dried over sodiumsulfate. Concentration in vacuo left a yellow oil, which was purified byflash column chromatography (60% ethyl acetate-hexanes) to give the4-methylphenyl sulfonamide derivative (1.2 mg, 93%) as a white solid.

EXAMPLE 30 7-Methylquinoline-2-carboxylic Acid Amide Derivative

A solution of the amine 4 (1.0 mg, 1.9 μmol, 1.0 equiv) indichloromethane (0.1 mL) was added to 7-methylquinoline-2-carboxylicacid (2.8 μmol, 1.5 equiv) and 1-hydroxybenzotriazole (0.44 mg, 3.2μmol, 1.7 equiv) in a vial at 23° C. The solution was stirred for 5 min,then N-cyclohexylcarbodiimide-N′-propyloxymethyl polystyrene (ArgonautTechnologies, 1.13 mmol/g, 3.4 mg, 3.8 μmol, 2.0 equiv) was added in oneportion at 23° C. The reaction mixture was stirred gently at 23° C.under an argon atmosphere for 16 h. PS-Trisamine (Argonaut Technologies,4.71 mmol/g, 2.0 mg, 9.5 μmol, 5.0 equiv) was then added in one portionand the reaction mixture was stirred for a further 2 h. The mixture wasfiltered through Celite (0.5 cm) and concentrated in vacuo to give the7-methylquinoline-2-carboxylic acid amide derivative (1.2 mg, 95%) as anoff-white solid.

¹H NMR (500 MHz, CDCl₃), δ8.18 (d, 1H, J=8.8, ArH), 8.07 (d, 1H, J=8.3,ArH), 7.93 (br t, 1H, J=5.8, NH), 7.73 (d, 1H, J=8.3, ArH), 7.62 (s, 1H,ArH), 7.43 (dd, 1H, J=8.3, 1.5, ArH), 5.72 (br s, 1H, ArOH), 5.42 (br s,1H, ArOH), 4.39 (t, 1H, J=3.9, CHCH₂NHCO), 4.19 (d, 1H, J=2.9, CHC≡N),4.16 (d, 1H, J=1.5, ArCHNCH₃), 3.81-3.77 (m, 4H, CH₂NHCO, ArOCH₃), 3.64(s, 3H, ArOCH₃), 3.58 (s, 3H, ArOCH₃), 3.51 (ddd, 1H, J=13.2, 4.4, 4.4,CH₂NHCO), 3.42 (br d, 1H, J=8.8, CHCHC≡N), 3.36 (s, 3H, ArOCH₃),3.32-3.25 (m, 2H, ArCHCHCH₂Ar, ArCHCHCH₂Ar), 3.07 (dd, 1H, J=18.1, 8.3,CH₂CHCHC≡N), 2.75 (d, 1H, J=18.1, CH₂CHCHC≡N), 2.59 (s, 3H, ArCH₃), 2.28(s, 3H, NCH₃), 2.23 (s, 3H, ArCH₃), 2.14 (dd, 1H, J=15.4, 11.5,ArCHCHCH₂Ar), 1.92 (s, 3H, ArCH₃); FTIR (neat film, cm⁻¹) 3383 (m, br,OH/NH), 2934 (m), 2228 (w, C≡N), 1668 (s, C≡O), 1463 (s); HRMS (ES)Calcd for C₃₉H₄₃N₅O₇ (MH)⁺: 694.3240, found: 694.3219.

EXAMPLE 31 Quinoxaline-2-carboxylic Acid Amide Derivative

¹H NMR (500 MHz, CDCl₃), δ9.47 (s, 1H, ArH), 8.14 (d, 1H, J=8.3, ArH),7.85-7.82 (m, 1H, ArH), 7.79-7.76 (m, 2H, ArH), 7.53 (dd, 1H, J=6.6,4.6, NH), 5.69 (s, 1H, ArOH), 5.35 (s, 1H, ArOH), 4.39 (t, 1H, J=3.2,CHCH₂NHCO), 4.18 (d, 1H, J=2.4, CHC≡N), 4.13 (d, 1H, J=1.9, ArCHNCH₃),3.88 (ddd, 1H, J=13.7, 7.6, 4.6, CH₂NHCO), 3.79 (s, 3H, ArOCH₃), 3.64(1H, ddd, J=13.7, 4.3, 3.0, CH₂NHCO), 3.61 (3H, s, ArOCH₃), 3.60 (3H, s,ArOCH₃), 3.44 (br d, 1H, J=8.6, CHCHC≡N), 3.29-3.25 (m, 2H, ArCHCHCH₂Ar,ArCHCHCH₂Ar), 3.25 (s, 3H, ArOCH₃), 3.04 (dd, 1H, J=18.5, 8.3,CH₂CHCHC≡N), 2.70 (d, 1H, J=18.5, CH₂CHCHC≡N), 2.25 (s, 3H, NCH₃), 2.23(s, 3H, ArCH₃), 2.12 (dd, 1H, J=16.1, 12.2, ArCHCHCH₂Ar), 1.88 (s, 3H,ArCH₃); FTIR (neat film, cm⁻¹) 3383 (m, br, OH/NH), 2933 (m), 2228 (w,C≡N), 1673 (s, C═O), 1463 (s); HRMS (ES) Calcd for C₃₇H₄₀N₆O₇ (MH)⁺:681.3036, found: 681.3062.

EXAMPLE 32 6-Chloroquinoline-2-carboxylic Acid Amide Derivative

¹H NMR (500 MHz, CDCl₃), δ8.14 (s, 2H, ArH), 7.85 (t, 1H, J=5.6, NH),7.82 (d, 1H, J=2.0, ArH), 7.74 (d, 1H, J=9.2, ArH), 7.67 (dd, 1H, J=9.3,2.4, ArH), 5.69 (s, 1H, ArOH), 5.45 (s, 1H, ArOH), 4.39 (t, 1H, J=3.7,CHCH₂NHCO), 4.17 (br s, 2H, CHC≡N, ArCHNCH₃), 3.76 (s, 3H, ArOCH₃), 3.72(ddd, 1H, J=13.7, 6.3, 4.4, CH₂NHCO), 3.64 (ddd, 1H, J=13.7, 5.3, 3.9,CH₂NHCO), 3.56 (s, 3H, ArOCH₃), 3.54 (s, 3H, ArOCH₃), 3.49 (s, 3H,ArOCH₃), 3.43 (br d, 1H, J=8.3, CHCHC≡N), 3.30-3.27 (m, 2H, ArCHCHCH₂Ar,ArCHCHCH₂Ar), 3.07 (dd, 1H, J=18.3, 8.3, CH₂CHCHC≡N), 2.69 (d, 1H,J=18.3, CH₂CHCHC≡N), 2.28 (s, 3H, NCH₃), 2.20 (s, 3H, ArCH₃), 2.16 (obsdd, 1H, J=16.2, 11.7, ArCHCHCH₂Ar), 1.95 (s, 3H, ArCH₃); FTIR (neatfilm, cm⁻¹) 3383 (m, br, OH/NH), 2923 (s), 2228 (w, C≡N), 1668 (s, C═O),1458 (s); HRMS (ES) Calcd for C₃₈H₄₀N₅O₇Cl (MH)⁺: 714.2694, found:714.2721.

EXAMPLE 33 Pentafluorobenzamide Derivative

¹H NMR (500 MHz, CDCl₃), δ5.66-5.65 (br m, 2H, ArOH, NH), 5.57 (s, 1H,ArOH), 4.27 (br m, 1H, CHCH₂NHCO), 4.17 (d, 1H, J=1.5, ArCHNCH₃),4.05-4.01 (m, 2H, CHC≡N, CH₂NHCO), 3.76 (s, 3H, ArOCH₃), 3.74 (s, 3H,ArOCH₃), 3.64 (s, 3H, ArOCH₃), 3.57 (s, 3H, ArOCH₃), 3.44 (br d, 1H,J=8.3, CHCHC≡N), 3.34-3.30 (m, 2H, ArCHCHCH₂Ar, CH₂NHCO), 3.25 (dd, 1H,J=16.2, 2.9, ArCHCHCH₂Ar), 3.05 (dd, 1H, J=18.5, 7.8, CH₂CHCHC≡N), 2.43(d, 1H, J=18.5, CH₂CHCHC≡N), 2.32 (s, 3H, NCH₃), 2.21 (s, 3H, ArCH₃),2.04 (s, 3H, ArCH₃), 1.95 (dd, 1H, J=16.2, 12.2, ArCHCHCH₂Ar); FTIR(neat film, cm⁻¹) 3363 (m, br, OH/NH), 2933 (m), 2228 (w, C≡N), 1678 (s,C═O), 1502 (s); HRMS (ES) Calcd for C₃₅H₃₅N₄O₇F₅ (MH)⁺: 719.2504, found:719.2479.

EXAMPLE 34 3-Methoxybenzamide Derivative

¹H NMR (500 MHz, CDCl₃), δ7.10 (br s, 1H, ArH), 7.07 (t, 1H, J=7.8,ArH), 6.91 (dd, 1H, J=8.3, 1.9, ArH), 6.19 (d, 1H, J=7.8, ArH), 6.00 (d,1H, J=5.9, NH), 5.69 (br s, 1H, ArOH), 5.64 (s, 1H, ArOH), 4.31 (br d,1H, J=2.4, CHCH₂NHCO), 4.23 (d, 1H, J=2.0, ArCHNCH₃), 4.13 (obs ddd, 1H,J=13.7, 8.3, 2.0, CH₂NHCO), 4.10 (d, 1H, J=2.4, CHC≡N), 3.79 (s, 3H,ArOCH₃), 3.75 (s, 3H, ArOCH₃), 3.74 (s, 3H, ArOCH₃), 3.48 (s, 3H,ArOCH₃), 3.45 (br d, 1H), J=8.8, CHCHC≡N), 3.42 (s, 3H, ArOCH₃),3.36-3.27 (m, 3H, ArCHCHCH₂Ar, CH₂NHCO (1H), ArCHCHCH₂Ar (1H)), 3.10(dd, 1H, J=18.6, 8.3, CH₂CHCHC≡N), 2.43 (d, 1H, J=18.6, CH₂CHCHC≡N),2.32 (s, 3H, NCH₃), 2.16 (s, 3H, ArCH₃), 2.08 (s, 3H, ArCH₃), 2.03 (dd,1H, J=16.1, 11.7, ArCHCHCH₂Ar); FTIR (neat film, cm⁻¹) 3395 (m, br,OH/NH), 2933 (m), 2226 (w, C≡N), 1651 (m, C═O), 1462 (s); HRMS (ES)Calcd for C₃₆H₄₂N₄O₈ (MH)⁺: 659.3081, found: 659.3081.

EXAMPLE 35 Hemiaminal Derivative

A solution of silver nitrate (15.9 mg, 93.6 μmol, 58.5 equiv) in amixture of water and acetonitrile (3:2, 500 μl) was added to thequinoline-2-carboxylic acid amide derivative (1.1 mg, 1.6 μmol, 1equiv). The reaction mixture was stirred at 23° C. in the dark for 4 h.A 1:1 mixture of saturated aqueous sodium bicarbonate and saturatedaqueous sodium chloride solution (2 mL, freshly prepared) was added andthe mixture was stirred for 5 min. A further 1:1 mixture of saturatedaqueous sodium bicarbonate and saturated aqueous sodium chloridesolution (20 mL, freshly prepared) was then added. The mixture wasextracted with methylene chloride (5×10 ml). The combined organicextract was dried over sodium sulfate and was then concentrated to leavea solid residue. The solid residue was purified by flash columnchromatography (4% methanol-methylene chloride) to provide thehemiaminal (0.8 mg, 74%) as a white solid.

¹H NMR (500 MHz, CDCl₃), δ8.22 (d, 1H, J=8.5, ArH), 8.12 (d, 1H, J=8.5,ArH), 8.04 (t, 1H, J=5.5, NH), 7.82 (d, 1H, J=8.5, ArH), 7.72 (m, 2H,ArH), 7.59 (ddd, 1H, J=8.5, 4.5, 1.5, ArH), 5.72 (br s, 1H, ArOH), 5.54(s, 1H, ArOH), 4.84 (t, 1H, J=4.0, CHCH₂NHCO), 4.60 (d, 1H, J=2.0,CHOH), 4.12 (dd, 1H, J=3.5, 1.5, CHCHOH), 3.78-3.68 (m, 4H, CH₂NHCO,ArOCH₃), 3.63 (1s, 3H, ArOCH₃), 3.47 (1s, 3H, ArOCH₃), 3.45 (1s, 3H,ArOCH₃). 3.38 (m, 1H, CH₂NHCO), 3.27 (m, 2H, ArCHCHCH₂Ar, ArCHCHCH₂Ar),3.01 (dd, 1H, J=16.5, 8.0, ArCHCHCH₂Ar), 2.60 (d, 2H, J=17, CH₂CHCHOH),2.27 (s, 3H, NCH₃), 2.21 (dd, 1H, J=16.5, 11.5, ArCHCHCH₂Ar), 2.17 (1s,3H, ArCH₃), 1.92 (1s, 3H, ArCH₃). FTIR (neat film), cm⁻¹ 3360, 2936,1667, 1415. HRMS (ES) Calcd for C₃₇H₄₃N₄O₈ (MH)⁺: 671.3081, Found:671.3083.

EXAMPLE 36 Methoxy Derivative

A solution of the quinoline-2-carboxylic acid amide derivative (0.8 mg,1.2 μmol, 1 equiv) in methanol (0.5 mL) was added, via cannula, tosilver tetrafluoroborate (22 mg, 113 μmol, 96 equiv) in an oven-driedglass vial. The reaction mixture was stirred at 23° C. in the dark for24 h. A 1:1 mixture of saturated aqueous sodium bicarbonate andsaturated aqueous sodium chloride solution (1 mL, freshly prepared) wasadded and the mixture was stirred for 5 min. A further 1:1 mixture ofsaturated aqueous sodium bicarbonate and saturated aqueous sodiumchloride solution (10 mL, freshly prepared) was then added. The mixturewas extracted with methylene chloride (4×10 mL). The combined organicextract was dried over sodium sulfate and was concentrated. The solidresidue was purified by flash column chromatography (4%methanol-methylene chloride) to provide the hemiaminal (0.45 mg, 57%) asa white solid and the methoxy derivative (0.16 mg, 19.8%) as a whitesolid.

¹H NMR of the methoxy derivative (500 MHz, CDCl₃), δ8.19 (d, 1H, J=8.0,ArH), 8.07 (d, 1H, J=8.0, ArH), 7.83 (m, 2H, NH, ArH), 7.76 (t, 1H,J=7.0, ArH), 7.62 (t, 1H, J=7.0, ArH), 7.53 (d, 1H, J=8.0, ArH), 5.60(br s, 1H, ArOH), 5.33 (s, 1H, ArOH), 4.64 (s, 1H, CHCH₂NHCO), 4.59 (d,1H, J=3.0, CHOCH₃), 4.18 (d, 1H, J=3.5, 1.5, CH₂NHCO), 4.06 (dd, 1H,J=14, 7.5, CHCHOH), 3.78 (1s, 3H, ArOCH₃), 3.69 (1s, 3H, ArOCH₃), 3.59(m, 1H, CH₂NHCO), 3.37 (m, 1H, ArCHCHCH₂Ar), 3.29 (m, 1H, ArCHCHCH₂Ar),3.16 (1s, 3H, ArOCH₃), 3.05 (1s, 3H, ArOCH₃), 2.99 (dd, 1H, J=18.5, 9,CH₂CHCHOCH₃), 2.88 (d, 1H, J=9, CH₂CHCHOCH₃), 2.52 (s, 3H, CHOCH₃), 2.36(d, J=18.5, ArCHCHCH₂Ar), 2.27 (s, 3H, NCH₃), 1.96 (ddd, J=18.5, 11,2.5, ArCHCHCH₂Ar), 1.76 (1s, 3H, ArCH₃), 1.74 (1s, 3H, ArCH₃). HRMS (ES)Calcd for C₃₈H₄₅N₄O₈ (MH)⁺: 685.3237, Found: 685.3259.

EXAMPLE 37 Reduced quinalascidin Analog

A solution of borane tetrahydrofuran complex in tetrahydrofuran (1.0 M,9.0 μL, 9 μmol, 5.0 equiv) was added to a solution of thequinoline-2-carboxylic acid amide derivative (1.2 mg, 1.8 μmol, 1 equiv)in tetrahydrofuran (0.2 mL). The resulting colorless solution wasstirred at 23° C. for 2.5 h. 1,4-Diazabicyclo[2.2.2]octane (DABCO, 6.1mg, 0.054 mmol, 30 equiv) and water (0.2 mL) were added to the reactionmixture in sequence. The resulting colorless solution was stirred at 23°C. for 20 h. The solution was diluted with dichloromethane (5 mL) andthe resulting mixture was dried over sodium sulfate and wasconcentrated. Purification of the white solid residue by flashchromatography (10% methanol-distilled ethyl acetate) afforded thereduced derivative (1.0 mg, 87%) as a white solid: R_(f) 0.16, 10%methanol-ethyl acetate; ¹H NMR (500 MHz, CD₃OD) δ8.17 (d, 1H, J=8.5 Hz,CHCHCCONH), 8.08 (d, 1H, J=8.5 Hz, CHCHCCONH), 8.05 (br t, 1H, CONH),7.80 (d, 1H, J=8.5 Hz, NCCH), 7.73 (ddd, 1H, J=7.5, 6.8, 1.5 Hz,NCCHCH), 7.70 (d, 1H, J=8.0 Hz, NCCHCHCHCH), 7.57 (ddd, 1H, J=8.0, 7.0,1.5 Hz, NCCHCHCH), 5.75 (br s, 1H, ArOH), 5.55 (s, 1H, ArOH), 4.20 (brs, 1H, ArCHNCH₃), 4.02 (br t, 1H, CHCH₂NHCO), 3.85 (br m, 1H,CHCH₂NHCO), 3.73 (s, 3H, ArOH), 3.67 (dt, 1H, J=13.0, 4.5 Hz,CHCH₂NHCO), 3.60 (s, 3H, ArOH), 3.44 (s, 3H, ArOH), 3.43 (s, 3H, ArOH),3.18-3.23 (m, 3H, ArCHCHCH₂Ar, ArCH₂CHCH₂N, ArCH₂CHCH₂N), 3.05 (dd, 1H,J=18.0, 7.5 Hz, ArCH₂CHCH₂N), 2.90-3.00 (m, 2H, ArCHCHCH₂Ar,ArCH₂CHCH₂N), 2.71 (d, 1H, J=20.0 Hz, ArCH₂CHCH₂N), 2.29 (s, 3H, NCH₃),2.23 (dd, 1H, J=15.5, 12.0 Hz, ArCHCHCH₂Ar), 2.13 (s, 3H, ArCH₃), 1.92(s, 3H, ArCH₃); FTIR (neat film), cm⁻¹ 3364 (br, NH, OH), 1668 (s, C═O);HRMS (TOF MS ES+) m/z calcd for C₃₇H₄₃N₄O₇ (M+H)⁺ 655.3132, found655.3123.

EXAMPLE 38 Synthesis of Labeled Analogs

As described generally above, certain of the inventive compounds canalso be modified to permit attachment of labeling reagents. For example,certain aryl and heteroaryl groups (and other groups) as definedgenerically herein, can be modified (by attachment of a linkerstructure, generally an aliphatic, heteroaliphatic, aryl, or heteroarylmoiety (or any combination thereof) which moiety is substituted orunsubstituted, cyclic or acyclic, branched or unbranched.Methyl ester 5:

A solution of trimethylsilyldiazomethane in hexanes (2.0M, 0.241 mL,0.484 mmol, 2.1 equiv) was added dropwise via syringe to a suspension ofacid 4 (52.0 mg, 0.231 mmol, 1 equiv) in a mixture of benzene (3.5 mL)and methanol (1.0 mL). The resulting red solution was stirred at 23° C.for 35 min. The solution was concentrated and the residual red oil waspurified by flash column chromatography (25→35% ethyl acetate-hexanes).Methyl ester 5 was obtained as a white solid (44.5 mg, 94%); abis-methylated biproduct (3.2 mg, 6%) was also isolated. Methyl ester 5:R_(f) 0.39, ethyl acetate; ¹H NMR (400 MHz, CDCl₃) δ8.22 (d, 1H, J=8.2Hz, CHCHCOH), 8.16 (d, 1H, J=8.4 Hz, CHCHCO₂CH₃), 8.13 (d, 1H, J=9.2 Hz,CHCHCO₂CH₃), 7.40 (dd, 1H, J=9.2, 2.8 Hz CHCHCOH), 7.17 (d, 1H, J=3.2Hz, CHCOH), 5.66 (br s, 1H, OH), 4.07 (s, 3H, OCH₃); ¹³C NMR (100 MHz,CD₃OD) δ166.9, 159.3, 145.5, 143.8, 137.0, 132.8, 132.1, 124.6, 122.1,109.1, 53.2; FTIR (neat film), cm¹ 3106 (br, OH), 1726 (m, C═O), 1228(s, Ar—OH); HMRS (CI) m/z calcd for C₁₁H₁₀NO₃ (M+H)⁺ 204.0661, found204.0665.Ether 6:

Diethyl azodicarboxylate (0.255 mL, 1.62 mmol, 4.0 equiv) was addeddropwise via syringe to an ice-cold solution of phenol 5 (82.2 mg, 0.405mmol, 1 equiv), N-Fmoc-1-amino-4-butanol (0.504 g, 1.62 mmol, 4.0equiv), and triphenylphosphine (0.424 g, 1.62 mmol, 4.0 equiv) intetrahydrofuran (2.7 mL). The resulting yellow solution was allowed towarm to 23° C. and was stirred for 19 h. The solution was concentrated.Purification of the residual yellow oil by flash column chromatography(50% ether-pentane→70% ether-pentane) afforded ether 6 as a white solid(126.8 mg, 63%): R_(f) 0.23, 50% ethyl acetate-hexanes; ¹H NMR (datagiven for major rotamer, 500 MHz, CDCl₃) δ8.19 (d, 1H, J=9.0 Hz, C₁₀H),8.13-8.16 (m, 2H, C₄H, C₅H), 7.76 (d, 2H, J=7.5 Hz, C₂₃H), 7.59 (d, 2H,J=7.5 Hz, C₂₀H), 7.38-7.43 (m, 3H, C₂₂H, C₉H), 7.30 (t, 2H, J=7.5 Hz,C₂₁H), 7.09 (d, 1H, J=2.0 Hz, C₇H), 4.86 (br m, 1H, CONH), 4.43 (d, 2H,J=7.0 Hz, CO₂CH₂), 4.21 (t, 1H, J=6.5 Hz, CO₂CH₂CH), 4.13 (t, 2H, J=5.8Hz, C₁₂H₂), 4.07 (s, 3H, OCH₃), 3.31 (q, 2H, J=6.3 Hz, C₁₅H₂), 1.91(quint, 2H, J=7.0 Hz, C₁₃H₂), 1.76 (quint, 2H, J=7.5 Hz, C₁₄H₂); ¹³C NMR(100 MHz, CDCl₃) δ166.1, 158.7, 156.5, 145.3, 143.9, 143.6, 141.3,135.6, 132.1, 130.8, 127.6, 127.0, 124.9, 123.6, 121.4, 119.9, 105.2,67.8, 66.5, 47.2, 40.5, 29.7, 26.7, 26.2; FTIR (neat film), cm⁻¹ 3334(br, NH), 1720 (br s, OC═O, NHC═O), 1227 (s, Ar—OCH₂).Carboxylic Acid 7:

A solution of methyl ester 6 (123 mg, 0.247 mmol) in a mixture of 3 Maq. HCl (0.5 mL) and p-dioxane (2.5 mL) was heated at reflux for 9 h.The solution was allowed to cool to 23° C., and was concentrated. Theresidual yellow solid was recrystallized from ethanol:water (2:1, 30mL). Upon cooling to −20° C., a white amorphous solid precipitated. Thesolid was collected by vacuum filtration and was washed with water (2mL), yielding acid 7 (106 mg, 82%) as a white powder: R_(f) 0.0, 50%ethyl acetate-hexanes; ¹H NMR (500 MHz, CD₃OD) δ8.36 (d, 1H, J=9.0 Hz,C₄H), 8.09-8.13 (m, 2H, C₃H, C₉H), 7.77 (d, 2H, J=7.5 Hz, C₂₂H), 7.62(d, 2H, J=7.5 Hz, C₂₀H), 7.49 (dd, 1H, J=9.5, 2.5 Hz, C₈H), 7.34-7.38(m, 3H, C₂₁H, C₆H), 7.27 (t, 2H, J=7.3 Hz, C₂₀H), 4.34 (d, 2H, J=6.5 Hz,CO₂CH₂), 4.18 (t, 2H, J=6.0 Hz, ArOCH₂), 4.16 (t, 1H, J=6.5 Hz,CO₂CH₂CH), 3.21 (t, 2H, J=6.5 Hz, CH₂NHCO₂), 1.88 (quint, 2H, J=7.0 Hz,ArOCH₂CH₂), 1.72 (quint, 2H, J=7.3 Hz, CH₂CH₂NHCO₂); ¹³C NMR (100 MHz,CD₃OD) δ165.7, 159.5, 157.0, 146.1, 145.2, 143.3, 142.1, 136.8, 131.9,131.7, 128.1, 127.6, 125.5, 124.2, 121.4, 120.6, 106.4, 68.6, 48.2,40.9, 30.3, 27.3, 26.9; FTIR (neat film), cm⁻¹ 3326 (br, CO₂H, NH), 1714(s, HOC═O, NHCO₂), 1227 (s, Ar—OCH₂); HRMS (TOF MS ES+) m/z calcd forC₂₉H₂₇N₂O₅ (M+H)⁺ 483.1920, found 483.1924.Amide 9:

A solution of primary amine 8 (13.8 mg, 26.3 μmol, 1.02 equiv) indichloromethane (0.75 mL) was added via cannula to a mixture of acid 7(12.5 mg, 25.9 μmol, 1 equiv) and 1-hydroxybenzotriazole (5.9 mg, 43.7μmol, 1.7 equiv) in an oven-dried glass vial. To the resultingsuspension was added PS-carbodiimide (Argonaut Technologies, 1.39mmol/g, 37.3 mg, 51.8 μmol, 2.0 equiv). The resulting suspension wasstirred gently at 23° C. for 19 h. The reaction mixture was directlypurified by chromatography on a pipette column of silica gel (2%methanol-dichloromethane). Amide 9 was obtained as a colorless oil (22.2mg, 87%): R_(f) 0.59, 10% methanol-dichloromethane; ¹H NMR (500 MHz,CDCl₃) δ8.06 (m, 2H, CHCHCCONH, CHCHCCONH), 7.90 (t, 1H, J=6.0 Hz,CONH), 7.77 (d, 2H, J=7.5 Hz, ArH), 7.69 (d, 1H, J=9.5 Hz, CHCHCOCH₂),7.60 (d, 2H, J=7.5 Hz, ArH), 7.40 (t, 2H, J=7.3 Hz, ArH), 7.35 (dd, 1H,J=9.5, 2.5 Hz, CHCHCOCH₂), 7.31 (t, 2H, J=7.5 Hz, ArH), 7.04 (d, 1H,J=2.5 Hz, CHCOCH₂), 5.71 (s, 1H, ArOH), 5.45 (s, 1H, ArOH), 4.85 (br m,1H, OCONH), 4.44 (d, 2H, J=7.0 Hz, CO₂CH₂), 4.37 (t, 1H, J=3.8 Hz,CHCH₂NHCO), 4.22 (t, 1H, J=7.0 Hz, CO₂CH₂CH), 4.17-4.20 (m, 2H, CHC≡N,ArCHNCH₃), 4.14-4.18 (m, 2H, ArOCH₂), 3.75 (s, 3H, ArOCH₃), 3.69-3.75(m, 1H, CH₂NHCO), 3.57 (s, 3H, ArOCH₃), 3.53 (s, 3H, ArOCH₃), 3.52-3.57(m, 1H, CH₂NHCO), 3.45 (s, 3H, ArOCH₃), 3.41 (d of m, 1H, J=7.0 Hz,CHCHC≡N), 3.32 (q, 2H, J=6.7 Hz, CH₂NHCO₂), 3.24-3.30 (m, 2H,ArCHCHCH₂Ar, ArCHCHCH₂Ar), 3.07 (dd, 1H, J=18.5, 8.0 Hz, CH₂CHCHC≡N),2.74 (d, 1H, J=18.5 Hz, CH₂CHCHC≡N), 2.27 (s, 3H, NCH₃), 2.19 (s, 3H,ArCH₃), 2.12 (dd, 1H, J=16.3, 12.3 Hz, ArCHCHCH₂Ar), 1.99 (s, 3H,ArCH₃), 1.88-1.94 (m, 2H, CH₂CH₂OAr), 1.76-1.80 (m, 2H, CH₂CH₂NHCO₂);FTIR (neat film), cm⁻¹ 3378 (br, NH, OH), 2233 (w, C≡N), 1714 (m,NHCO₂), 1668 (m, NHC═O); HRMS (TOF MS ES+) m/z calcd for C₅₇H₆₁N₆O₁₀(M+H)⁺: 989.4449, found 989.4404.Primary Amine 10:

1,8-Diazabicyclo[5.4.0]undec-7-ene (DBU, 0.5 μL, 3.6 μmol, 1.5 equiv)was added to a solution of carbamate 9 (2.4 mg, 2.4 μmol, 1 equiv) indichloromethane (0.1 mL). The resulting solution was stirred for 30 min.Purification of the crude reaction mixture by column chromatography on apipette column (4% methanol-dichloromethane→10%methanol-dichloromethane→10% methanol in 98:2 dichloromethane:ammoniumhydroxide), furnished primary amine 10 as a white solid (1.7 mg, 92%):R_(f) 0.03, 10% methanol-ichloromethane; ¹H NMR (400 MHz, CDCl₃) δ8.06(m, 2H, CHCHCCONH, CHCHCCONH), 7.90 (t, 1H, J=5.2 Hz, CONH), 7.69 (d,1H, J=9.2 Hz, CHCHCOCH₂), 7.35 (dd, 1H, J=9.2, 2.8 Hz, CHCHCOCH₂), 7.03(d, 1H, J=2.8 Hz, CHCOCH₂), 4.37 (t, 1H, J=3.6 Hz, CHCH₂NHCO), 4.15-4.18(m, 2H, CHC≡N, ArCHNCH₃), 4.11 (t, 1H, J=6.4 Hz, ArOCH₂), 4.11 (t, 1H,J=6.4 Hz, ArOCH₂), 3.75 (s, 3H, ArOCH₃), 3.69-3.75 (m, 1H, CH₂NHCO),3.58 (s, 3H, ArOCH₃), 3.53-3.58 (m, 1H, CH₂NHCO), 3.53 (s, 3H, ArOCH₃),3.46 (s, 3H, ArOCH₃), 3.40-3.43 (m, 1H, CHCHC≡N), 3.23-3.30 (m, 2H,ArCHCHCH₂Ar, ArCHCHCH₂Ar), 3.08 (dd, 1H, J=18.4, 8.4 Hz, CH₂CHCHC≡N),2.83 (br m, 2H, CH₂NH₂), 2.74 (d, 1H, J=18.4 Hz, CH₂CHCHC≡N), 2.27 (s,3H, NCH₃), 2.19 (s, 3H, ArCH₃), 2.12 (dd, 1H, J=16.2, 11.8 Hz,ArCHCHCH₂Ar), 1.99 (s, 3H, ArCH₃), 1.88-1.96 (m, 2H, CH₂CH₂OAr),1.65-1.74 (m, 2H, CH₂CH₂NH₂); FTIR (neat film), cm⁻¹ 3378 (br, NH, OH),2248 (w, C≡N), 1668 (m, NHC═O); HRMS (TOF MS ES+) m/z calcd forC₄₂H₅₁N₆O₈ (M+H)⁺: 767.3768, found: 767.3759.Biotinylated quinalascidin Analog 1:

A suspension of primary amine 10 (0.8 mg, 1.0 μmol, 1 equiv) andN-hydroxy-succinimidyl ester 11 (Molecular Probes, 0.71 mg, 1.3 μmol,1.3 equiv) in tetrahydrofuran (0.1 mL) in a glass vial was stirred at23° C. for 14 h. The mixture was concentrated and the residual whitesolid was purified by column chromatography on a pipette column (5%methanol-dichloromethane→10% methanol-dichloromethane→10% methanol in98:2 dichloromethane:ammonium hydroxide). Fractions containing thebiotinylated product 1 were contaminated by remaining starting amine 10.Further purification by chromatography on Sephadex LH-20 (11 cm×1.3 cm,methanol, gravity) yielded 1 as a colorless oil (0.67 mg, 55%). Materialused in isolation of quinalascidin-binding proteins by affinityprecipitation was further purified by HPLC (Beckman Ultrasphere ODSreverse phase column, 10 mm×25 mm, flow rate 2.0 mL/min, isocraticelution with 40% acetonitrile-water, retention time 29 min): R_(f) 0.45,20% methanol-dichloromethane; ¹H NMR (600 MHz, CD₃OD) δ8.18 (d, 1H,J=8.4 Hz, CHCHCCONH), 7.85 (d, 1H, J=9.0 Hz, CHCHCCONH), 7.56 (d, 1H,J=9.0 Hz, CHCHCOCH₂), 7.40 (dd, 1H, J=9.0, 2.4 Hz, CHCHCOCH₂), 7.23 (d,1H, J=2.4 Hz, CHCOCH₂), 4.44-4.48 (m, 2H, CHC≡N, SCH₂CHNH), 4.38 (br t,1H, CHCH₂NHCO), 4.27 (dd, 1H, J=7.8, 4.0 Hz, SCHCHNH), 4.21 (br d, 1H,ArCHNCH₃), 4.13-4.17 (m, 2H, ArOCH₂), 3.94 (dd, 1H, J=13.8, 3.0 Hz,CHCH₂NHCO), 3.65 (s, 3H, ArOCH₃), 3.60-3.66 (m, 1H, CHCH₂NHCO), 3.43 (s,3H, ArOCH₃), 3.41-3.46 (m, 1H, CHCHC≡N), 3.39 (s, 3H, ArOCH₃), 3.38 (s,3H, ArOCH₃), 3.28-3.32 (obscured by solvent peak, 1H, ArCHCHCH₂Ar), 3.27(t, 2H, J=7.2 Hz, ArOCH₂CH₂CH₂CH₂NHCO), 3.14 (t, 4H, J=7.2 Hz,CH₂CH₂NHCO, CH₂CH₂NHCO), 3.13-3.18 (obscured m, SCH), 3.10 (d of m, 1H,J=11.4 Hz, ArCHCHCH₂Ar), 3.03 (dd, 1H, J=18.6, 8.4 Hz, CH₂CHCHC≡N), 2.89(dd, 1H, J=12.9, 5.1 Hz, SCH₂), 2.68 (d, 1H, J=12.6 Hz, SCH₂), 2.61 (d,1H, J=18.6 Hz, CH₂CHCHC≡N), 2.27 (dd, 1H, J=15.9, 11.7 Hz, ArCHCHCH₂Ar),2.22 (s, 3H, NCH₃), 2.14-2.22 (m, 6H, CH₂CO, CH₂CO, CH₂CO), 2.11 (s, 3H,ArCH₃), 1.86-1.92 (m, 2H, CH₂CH₂OAr), 1.85 (s, 3H, ArCH₃), 1.70-1.76 (m,2H, ArOCH₂CH₂CH₂), 1.26-1.70 (m, 18 H, COCH₂CH₂CH₂CH₂CH₂NH (both),CH₂CH₂CH₂CHS); LRMS (TOF MS ES+) m/z calcd for C₆₅H₉₁N₁₀O₁₂S (M+H)⁺1220, found 1220.Fluorescein-Labelled quinalascidin Analog 2:

A solution of primary amine 10 (0.695 mg, 0.91 μmol, 1 equiv) andN-hydroxy-succinimidyl ester 12 (Molecular Probes, 0.560 mg, 0.96 μmol,1.05 equiv) in tetrahydrofuran (0.2 mL) was stirred in the dark at 23°C. for 71 h. The yellow solution was concentrated. The solid residue waspurified by HPLC (Beckman Ultrasphere ODS reverse phase column, 10 mm×25mm, flow rate 2.0 mL/min, gradient elution from 5% to 60% acetonitrilein water over 40 min, injected crude product as a solution in 50%acetonitrile-water (200 μL), retention time 36 min). Fractionscontaining the desired product were pooled and were concentrated toremove acetonitrile. The residual aqueous solution was lyophilized,yielding fluorescein-labelled analog 2 (0.305 mg, 27%) as an orangesolid: R_(f) 0.49, 20% methanol-dichloromethane; ¹H NMR (600 MHz, CD₃OD)δ8.44 (br s, 1H, NHCOCCHCO₂), 8.16 (d, 1H, J=8.4 Hz, NCCHCH), 8.03 (brm, 1H, CO₂CCCHCHCCONH), 7.83 (d, 1H, J=9.0 Hz, NCCHCH), 7.54 (d, 1H,J=8.4 Hz, CHCHCOCH₂), 7.38 (dd, 1H, J=9.0, 2.4 Hz, CHCHCOCH₂), 7.29 (d,1H, J=7.8 Hz, CO₂CCCHCHCCONH), 7.20 (d, 1H, J=3.0 Hz, CHCOCH₂),6.58-6.67 (br m, 6H, CHCHCOHCH, CHCHCOHCH), 4.45 (d, 1H, J=2.4 Hz,CHC≡N), 4.37 (br s, 1H, CHCH₂NHCO), 4.19-4.21 (m, 1H, ArCHNCH₃),4.11-4.14 (m, 2H, ArOCH₂), 3.94 (dd, 1H, J=13.2, 3.0 Hz, CHCH₂NHCO),3.64 (s, 3H, ArOCH₃), 3.48 (dd, 1H, J=13.5, 3.3 Hz, CHCH₂NHCO), 3.44 (t,2H, J=7.2 Hz, CH₂NHCO), 3.42 (s, 3H, ArOCH₃), 3.38 (s, 3H, ArOCH₃), 3.37(s, 3H, ArOCH₃), 3.28 (t, 2H, J=7.2 Hz, CH₂NHCO), 3.09 (d of m, 1H,J=11.4 Hz, ArCHCHCH₂Ar), 3.02 (dd, 1H, J=18.6, 7.8 Hz, CH₂CHCHC≡N), 2.60(d, 1H, J=18.6 Hz, CH₂CHCHC≡N), 2.27 (dd, 1H, J=15.6, 11.4 Hz,ArCHCHCH₂Ar), 2.24 (t, 2H, J=7.2 Hz, CH₂CONH), 2.22 (s, 3H, NCH₃), 2.10(s, 3H, ArCH₃), 1.85-1.90 (m, 2H, CH₂CH₂OAr), 1.83 (s, 3H, ArCH₃),1.66-1.76 (m, 4H, CH₂CH₂NHCO, CH₂CH₂NHCO); LRMS (TOF MS ES+) m/z calcdfor C₆₉H₇₂N₇O₁₅ (M+H)⁺ 1239, found 1239.

IV. In vitro Activity:

a) Experimentals:

Cells and Cell Culture Conditions

The two cell lines used, A375 malignant melanoma and A-549 lungcarcinoma, were purchased from American Type Culture Collection. Thesecells were cultured at 37° C. in a humidified atmosphere of 5% CO₂ inDulbecco's Modified Eagle Medium (DMEM) supplemented with 10% fetalbovine serum (FBS), 0.1% glutamine and 0.1% penicillin-streptomycin.DMEM, FBS, glutamine and penicillin-streptomycin were purchased fromLife Technologies (Grand Island, N.Y.).

Cell Growth Inhibition Assay

Exponentially growing cells were seeded at ˜3000 (50 μl) cells per wellin 96-well flat-bottomed microtiter plates and then incubated at 37° C.in a humidified atmosphere of 5% CO₂/95% air for 24 hr. An analogue wasdissolved in dimethyl sulfoxide to give a concentration of 0.6 mg/ml,which was further diluted with the culture medium containing 10% fetalbovine serum. Nine three-fold dilutions were prepared with the maximumconcentration being 1/300 of the original DMSO solution (i.e. 2000ng/ml). This procedure was repeated for each analogue. Fifty microlitersof the obtained dilutions were each transferred into the well of theabove described culture plate. The cell culture was then placed backinto the incubator at 37° C. under the 5% CO₂ atmosphere for 72 hr. Cellproliferation was quantified by using the CellTiter 96 AQ_(ueous) Assay(Promega). In this assay a mixture solution (20 μl) of MTS [Owen'sreagent:3-(4,5-dimethylthiazol-2-yl)-5-(3-carbomethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazoliuminner salt] and phenazine methosulfate (PMS) was added to each well. Theresulting mixtures were further incubated for 2.5 hr. The absorbance wasmeasured with a microplate reader at a test wavelength of 540 nm and areference wavelength of 655 nm to serve as an index of the number ofviable cells. The inhibitory ratio of the test compound was calculatedaccording to the following formula: inhibition ratio (%)=100×(C−T)/C,where T is an absorbance of the well containing a test compound and C isan absorbance of the well containing no test compound. The IC₅₀ wascalculated by the least squares method.

b) Exemplary Data: TABLE 1 IC₅₀, nM⁸ N = A375 A549 Saframycin A (1) 5.3133

4.5 160

52 1400

330 1100

260 700 N—(Pyr)₂ >1400 >1400

370 1100

13 290

34 1200

2.4 39

3.9 50

R = NO₂ 3.6 91 R = CH₃ 1.9 37 R = Br 1.7 25

12 395

14 390

8.5 100

3.3 40

4.4 100

2.7 31

2.9 44

43 390

2.5 32

23 320

9.2 25

R = OH 11 91 R = OCH₃ 3.5 6.4 R = CH₃ 2.2 4.4 R = H (11) 1.3 4.4

1.4 4.6

3.4 96

37 890

81 860

R = Phenyl 34 330 R = I 23 790 R = Cl 11 340 R = Me 7.6 350 R = H 4.1 91R = F 2.5 37 R = OH (7) 1.4 14

1.2 11.3

1.2 6.5

4.1 99

76 1200

85 1100

2.5 26

9.6 110

96 1100

310 >1400

1.7 9.2

>510 >510

R = OH 29 120 R = OBu 4.7 13 R = CH₃ 2.5 4.0 R = OCH₃ (13) 2.0 3.5 R =Cl (14) 1.5 4.1

7.5 42

13 74

1.2 4.7

3.6 78

8.3 200

34 42 975 1100(N = R₁ in generic structure, where m is 1)

TABLE 2 IC₅₀ (nM) Cell line A375-Melanoma A549-Lung

35 176

116 1400

29 118 Biotinylated derivative 210 990 Fluorescein derivative 18 590

TABLE 3

IC₅₀ (nM) Cell line X A375-Melanoma A549-Lung OH 0.9 13 OCH₃ 120 1100H >9000 >9000V. In vivo Activity:

Although a variety of methods can be utilized, one exemplary method bywhich the in vivo activity of the inventive compounds is determined isby subcutaneously transplanting a desired tumor mass in mice. Drugtreatment is then initiated when tumor mass reaches approximately 100mm³ after transplantation of the tumor mass. A suitable composition, asdescribed in more detail above, is then administered to the mice,preferably in saline and also preferably administered once a day atdoses of 5, 10 and 25 mg/kg, although it will be appreciated that otherdoses can also be administered. Body weight and tumor size are thenmeasured daily and changes in percent ratio to initial values areplotted. In cases where the transplanted tumor ulcerates, the weightloss exceeds 25-30% of control weight loss, the tumor weight reaches 10%of the body weight of the cancer-bearing mouse, or the cancer-bearingmouse is dying, the animal is sacrificed in accordance with guidelinesfor animal welfare.

1-75. (canceled)
 76. A method for the synthesis of a compound having theformula (I):

wherein R₁ is NR_(A)R_(B), —OR_(A), —SR_(A), —C(═O)R_(A), —C(═S)R_(A),—S(O)₂R_(A), or an aliphatic, heteroaliphatic, aryl, heteroaryl,(aliphatic)aryl, (aliphatic)heteroaryl, (heteroaliphatic)aryl, or(heteroaliphatic)heteroaryl moiety, wherein each occurrence of R_(A) andR_(B) is independently hydrogen, —(C═O)R_(C), —NHR_(C), —(SO₂)R_(C),—OR_(C), or an aliphatic, heteroaliphatic, aryl, or heteroaryl moiety,or R_(A) and R_(B), when taken together form an aryl, heteroaryl,cycloaliphatic, or cycloheteroaliphatic moiety, wherein each occurrenceof R_(C) is independently hydrogen, —OR_(D), —SR_(D), —NHR_(D),—(C═O)R_(D), or an aliphatic, heteroaliphatic, aryl, or heteroarylmoiety, wherein each occurrence of R_(D) is independently hydrogen, aprotecting group, or an aliphatic, heteroaliphatic, aryl, heteroaryl,acyl, alkoxy, aryloxy, alkylthio, arylthio, heteroaryloxy, orheteroarylthio moiety; wherein R₂ is hydrogen, —OR_(E), ═O, —C(═O)R_(E),—CO₂R_(E), —CN, —SCN, halogen, —SR_(E), —SOR_(E), —SO₂R_(E), —NO₂,—N(R_(E))₂, —NHC(O)R_(E), or an aliphatic, heteroaliphatic, aryl, orheteroaryl moiety, wherein each occurrence of R_(E) is independentlyhydrogen, a protecting group, or an aliphatic, heteroaliphatic, aryl,heteroaryl, acyl, alkoxy, aryloxy, alkylthio, arylthio, heteroaryloxy,or heteroarylthio moiety; wherein R₃ is hydrogen, a nitrogen protectinggroup, —COOR_(F), —COR_(F), —CN, or an aliphatic, heteroaliphatic, aryl,or heteroaryl moiety, wherein each occurrence of R_(F) is independentlyhydrogen, a protecting group, or an aliphatic, heteroaliphatic, aryl,heteroaryl, alkoxy, aryloxy, alkylthio, arylthio, heteroaryloxy, orheteroarylthio moiety; wherein R₄ and R₆ are each independentlyhydrogen, or an aliphatic, heteroaliphatic, aryl, heteroaryl, acyl,alkoxy, aryloxy, alkylthio, arylthio, heteroaryloxy, or heteroarylthiomoiety; wherein R₅ and R₇ are each independently hydrogen, —OR_(G),—C(═O)R_(G), —CO₂R_(G), —CN, —SCN, halogen, —SR_(G), —SOR_(G),—SO₂R_(G), —NO₂, —N(R_(G))₂, —NHC(O)R_(G), or an aliphatic,heteroaliphatic, aryl or heteroaryl moiety, wherein each occurrence ofR_(G) is independently hydrogen, a protecting group, or an aliphatic,heteroaliphatic, aryl, heteroaryl, acyl, alkoxy, aryloxy, alkylthio,arylthio, heteroaryloxy, or heteroarylthio moiety; wherein R₈ ishydrogen, alkyl, —OH, protected hydroxyl, ═O, —CN, —SCN, halogen, —SH,protected thio, alkoxy, thioalkyl, amino, protected amino, oralkylamino; wherein m is 0-5; wherein X₁, X₂, X₃ and X₄ are eachindependently hydrogen, —OR_(H), ═O, —C(═O)R_(H), —CO₂R_(H), —CN, —SCN,halogen, —SR_(H), —SOR_(H), —SO₂R_(H), —NO₂, —N(R_(H))₂, —NHC(O)R_(H),or an aliphatic, heteroaliphatic, aryl, or heteroaryl moiety, whereineach occurrence of R_(H) is independently hydrogen, a protecting group,or an aliphatic, heteroaliphatic, aryl, heteroaryl, acyl, alkoxy,aryloxy, alkylthio, arylthio, heteroaryloxy, or heteroarylthio moiety;or wherein X₁ and R₇ taken together comprise a heterocyclic moiety;whereby if at least either X₁ and X₂ or X₃ and X₄ are doubly bonded tothe 6-membered ring, then the dotted bonds in either or both of the6-membered rings represent two single bonds and one double bond, and aquinone moiety is generated, or if at least either X₁ and X₂ or X₃ andX₄ are singly bonded to the 6-membered ring, then the dotted bonds ineither or both of the 6-membered rings represent two double bonds andone single bond, and a hydroquinone moiety is generated; whereby each ofthe foregoing aliphatic, heteroaliphatic and alkyl moieties mayindependently be substituted or unsubstituted, branched or unbranched,or cyclic or acyclic, and each of the foregoing aryl or heteroarylmoieties may independently be substituted or unsubstituted; wherein saidmethod comprises: (a) providing a compound of formula (XV)

(b) reacting said compound of formula (XV) under suitable conditions togenerate a compound of formula (I):

wherein X₁-X₄, R₁-R₈, and m are as described above, and wherein the stepof providing a compound of formula (XV) further comprises: (1) reactinga first N-protected and a second C-protected α-amino aldehyde precursorhaving the structures:

under suitable conditions to generate a tetrahydroisoquinoline corehaving the structure (IX):

(2) optionally reacting said tetrahydroisoquinoline core under suitableconditions to diversify R₃; (3) reacting a third aldehyde precursorhaving the structure: R₉(CH₂)_(m)CHO, with said tetrahydroisoquinolinecore structure (XIV) under suitable conditions to generate a trimer ofaldehydes having the structure:

(4) reacting said trimer of aldehydes under suitable conditions togenerate a compound of structure (XV), wherein P₁ is hydrogen or anitrogen protecting group; X₅ and X₆ taken together represent a carbonprotecting group, optionally substituted with a solid support unit; andR₉ is NR_(L)R_(M), —OR_(L), —SR_(L), —C(═O)R_(L), —C(═S)R_(L),—S(O)₂R_(L), or an aliphatic, heteroaliphatic, aryl, heteroaryl,(aliphatic)aryl, (aliphatic)heteroaryl, (heteroaliphatic)aryl, or(heteroaliphatic)heteroaryl moiety, wherein each occurrence of R_(L) andR_(M) is independently hydrogen, —(C═O)R_(N), —NHR_(N), —(SO₂)R_(N),—OR_(N), or an aliphatic, heteroaliphatic, aryl, or heteroaryl moiety,or R_(L) and R_(M), when taken together form an aryl, heteroaryl,cycloaliphatic, or cycloheteroaliphatic moiety, wherein each occurrenceof R_(N) is independently hydrogen, —OR_(P), —SR_(P), —NHR_(P),—(C═O)R_(P), or an aliphatic, heteroaliphatic, aryl, or heteroarylmoiety, wherein each occurrence of R_(P) is independently hydrogen, aprotecting group, or an aliphatic, heteroaliphatic, aryl, heteroaryl,acyl, alkoxy, aryloxy, alkylthio, arylthio, heteroaryloxy, orheteroarylthio moiety.
 77. The method of claim 76, wherein for theintermediates (XIV) and (XV) R₉ is —NHP₂, P₂ is a nitrogen protectinggroup, and the intermediates have the structures (XIVa) and (XVa):


78. The method of claim 76, wherein R₉(CH₂)_(m)CHO is(aliphatic)(C═O)(CH₂)_(m)CHO, (heteroaliphatic)(C═O) (CH₂)_(m)CHO,(aliphatic)(CH₂)_(m)CHO, (heteroaliphatic)(CH₂)_(m)CHO,aryl(aliphatic)(CH₂)_(m)CHO, aryl(heteroaliphatic)(CH₂)_(m)CHO,-heteroaryl(aliphatic)(CH₂)_(m)CHO, orheteroaryl(heteroaliphatic)(CH₂)_(m)CHO, wherein each of the aliphatic,and heteroaliphatic moieties is independently cyclic or acyclic, linearor branched, or substituted or unsubstituted and wherein the aryl andheteroaryl moieties are independently substituted or unsubstituted. 79.The method of claim 76, wherein R₉(CH₂)_(m)CHO is CH₃(CH₂)₁₋₆CHO;(protecting group)O(CH₂)₁₋₆CHO; (protecting group)NH(CH₂)₁₋₆CHO;(protecting group)S(CH₂)₁₋₆CHO; (alkyl)O(C═O)CHO; (aryl)(alkenyl)CHO;(heteroaryl)(alkenyl)CHO; (aryl)CHO; or (heteroaryl)CHO, wherein each ofthe aliphatic, and heteroaliphatic moieties is independently cyclic oracyclic, linear or branched, or substituted or unsubstituted and whereinthe aryl and heteroaryl moieties are independently substituted orunsubstituted.
 80. The method of claim 76, wherein X₅ is CN and X₆ is aheterocyclic moiety optionally substituted with a solid support unit.81. The method of claim 76, wherein the alkaloid structure (I) generatedis that of saframycin A.
 82. The method of claim 76, wherein the methodis stereoselective and the alkaloid structure (I) generated is that of−(−) saframycin A.
 83. The method of claim 76, wherein the compound offormula (I) has the structure of formula (Ia):


84. The method of claim 76, wherein the compound of formula (I) has thestructure of formula (II):


85. The method of claim 76, wherein the compound of formula (I) has thestructure of formula (III):


86. The method of claim 76, wherein the compound of formula (I) has thestructure of formula (IV):


87. The method of claim 76, wherein the compound of formula (I) has thestructure of formula (V):


88. The method of claim 76, wherein the compound of formula (I) has thestructure of formula (VI):


89. The method of claim 76, wherein the compound of formula (I) has thestructure of formula (VII):


90. The method of claim 76, wherein the compound of formula (I) has thestructure of formula (VIII):


91. The method of claim 76, wherein the compound of formula (I) has thestructure of formula (IX):

wherein R₁ is a substituted or unsubstituted, cyclic or acyclic,branched or unbranched aliphatic or heteroaliphatic moiety, or is asubstituted or unsubstituted aryl or heteroaryl moiety.
 92. The methodof claim 76, wherein the compound of formula (I) has the structure offormula (X):

wherein R₁ is a substituted or unsubstituted, cyclic or acyclic,branched or unbranched aliphatic or heteroaliphatic moiety, or is asubstituted or unsubstituted aryl or heteroaryl moiety.
 93. The methodof claim 76, wherein the compound of formula (I) has one or more of thefollowing limitations: when m is 1, R₁ excludes any one or more of thefollowing groups: —NH(protecting group), —NH₂, —NHCOCOMe,—NHCOC(Me)(OMe)(OMe), —NHCOCH(NH₂)CH₃,—NHCOCH(NH(acyl))CH₃—NHCOCH(NH₂)Ac, or NHCOCH(NHCOOBn)(Me);—O(C═O)C(CH₃)═C(CH₃)H; —OH, —O(protecting group), —O(COCH₃),—O(C═O)CH₂CH₃; or when m is 1; when X₁, X₂, X₃ and X₄ are each ═O; whenR₂ is —CN or —OH; when R₄ and R₆ are each —CH₃; when R₅ and R₇ are each—OCH₃; when R₈ is H; and R₁ is —NH(C═O)R_(C), then R_(C) is not—CH(NR_(W)R_(Y))(CH₂R_(Z)) where R_(W) and R_(Y) are each independentlyhydrogen or C₁₋₇ alkyl, aryl(C₁₋₄)alkyl, (C₁₋₄)alkylaryl, a substitutedsulfonyl (—S(O)₂—) group, or a substituted acyl group, and where R_(Z)is hydrogen or C₁₋₄ alkyl; or when m is 1; when X₁, X₂, X₃ and X₄ areeach ═O; when R₂ is —CN; when R₄ and R₆ are each —CH₃; when R₅ and R₇are each —OCH₃; when R₈ is H; and R₁ is —NH(C═O)R_(C), then R_(C) is not—C(OH)(Me)CH₂(C═O)Me; or when m is 1 and when R₂ is H; and R₁ is—NH(C═O)R_(C), then R_(C) is not —CH(Me)NH(C═O)O(CH₂)Ph; or when m is 0;R₂ is H; X₃ is H; and R₁ is —C(═O)R_(A), then R_(A) is not —O(alkyl); orwhen R₂ is H; m is 1; and R₁ is —OR_(A), then R_(A) is not —C(═O)R_(C),or S(O)₂R_(C), wherein R_(C) is an alkyl moiety.
 94. The method of claim76, wherein m is 0 or
 1. 95. The method of claim 76, wherein wherein R₂is CN, —SCN, ═O, OH, protected hydroxyl, H, or alkoxy.
 96. The method ofclaim 76, wherein R₂ is hydrogen, hydroxyl, —CN or —SCN.
 97. The methodof claim 76, wherein R₈ is hydrogen.
 98. The method of claim 76, whereinX₁, X₂, X₃, and X₄ are each independently alkoxy, OH, protectedhydroxyl, or ═O.
 99. The method of claim 76, wherein R₂ is CN, —SCN, ═O,OH, protected hydroxyl, H, or alkoxy; R₃ is hydrogen, a nitrogenprotecting group, —CN, aliphatic, or aryl; R₄ and R₆ are each alkyl; R₅and R₇ are each alkyloxy or thioalkyl; R₈ is hydrogen, alkyl, —OH,protected hydroxyl, ═O, CN, halogen, SH, alkoxy, thioalkyl, amino, oralkylamino; and X₁, X₂, X₃, and X₄ are each independently alkoxy, OH or═O.
 100. The method of claim 76, wherein R₂ is —CN, —SCN, —OH, protectedhydroxyl, H, or alkoxy; R₃ is hydrogen, a nitrogen protecting group,aliphatic, or aryl; R₄ and R₆ are each alkyl; R₅ and R₇ are eachalkyloxy or thioalkyl; X₁ and X₄ are each —OH; R₈ is hydrogen, alkyl,OH, protected hydroxyl, ═O, CN, halogen, SH, alkoxy, thioalkyl, amino,or alkylamino; and X₂ and X₃ are each alkyloxy or thioalkyl.
 101. Themethod of claim 76, wherein X₁ is OH, X₂ is OCH₃, X₃ is OCH₃, X₄ is OH,R₂ is CN, H or OH, R₃ is Me, R₄ is CH₃, R₅ is OCH₃, R₆ is CH₃, R₇ isOCH₃, and R₈ is H.
 102. The method of claim 76, wherein R₁ is OR_(A),SR_(A), or NR_(A)R_(B), wherein R_(A) and R_(B) are each independentlyhydrogen, —(C═O)R_(C) or an aliphatic, heteroaliphatic, aryl, orheteroaryl moiety, wherein R_(C) is —(C═O)R_(D), or an aliphatic,heteroaliphatic, aryl or heteroaryl moiety, and wherein R_(D) is analiphatic, heteroaliphatic, aryl, or heteroaryl moiety, or wherein R_(A)and R_(B), taken together, form a heterocyclic moiety, whereby each ofsaid aliphatic and heteroaliphatic moieties is independently substitutedor unsubstituted, branched or unbranched, or cyclic or acyclic, and eachof said aryl, heteroaryl and heterocyclic moieties is independentlysubstituted or unsubstituted.
 103. The method of claim 76, wherein R₁ isOR_(A), SR_(A), or NR_(A)R_(B), wherein R_(A) and R_(B) are eachindependently hydrogen, —(C═O)R_(C), or an aryl, (aliphatic)aryl,(heteroaliphatic)aryl, heteroaryl, (aliphatic)heteroaryl, or(heteroaliphatic)heteroaryl moiety, wherein R_(C) is an aryl,(aliphatic)aryl, (heteroaliphatic)aryl, heteroaryl,(aliphatic)heteroaryl, or (heteroaliphatic)heteroaryl moiety, or whereinR_(A) and R_(B) taken together form a heterocyclic moiety, whereby eachof said aliphatic and heteroaliphatic moieties is independentlysubstituted or unsubstituted, branched or unbranched, or cyclic oracyclic, and each of said aryl, heteroaryl and heterocyclic moieties isindependently substituted or unsubstituted.
 104. The method of claim 76,wherein R₁ is —NR_(A)C(═O)R_(C), wherein R_(A) is hydrogen or loweralkyl, and R_(C) is a substituted or unsubstituted, branched orunbranched, cyclic or acyclic aliphatic or heteroaliphatic moiety, or asubstituted or unsubstituted aryl or heteroaryl moiety, or wherein R_(A)and R_(C) taken together form a heterocyclic or heteroaryl moiety. 105.The method of claim 76, wherein R₁ is NR_(A)C(═O)R_(C), wherein R_(A) ishydrogen or lower alkyl, and R_(C) is an aryl, (aliphatic)aryl,(aliphatic)heteroaryl, heteroaryl, (heteroaliphatic)aryl, or(heteroaliphatic)heteroaryl moiety, or wherein R_(A) and R_(C) takentogether form a heterocyclic or heteroaryl moiety; whereby each of saidaliphatic and heteroaliphatic moieties is independently substituted orunsubstituted, branched or unbranched, or cyclic or acyclic, and each ofsaid aryl, heteroaryl and heterocyclic moieties is independentlysubstituted or unsubstituted.
 106. The method of claim 76, wherein R₁ isa substituted or unsubstituted, branched or unbranched, cyclic oracyclic aliphatic or heteroaliphatic moiety, or a substituted orunsubstituted aryl or heteroaryl moiety.
 107. The method of claim 76,wherein R₁ is an aryl, (aliphatic)aryl, (aliphatic)heteroaryl,heteroaryl, (heteroaliphatic)aryl, or (heteroaliphatic)heteroarylmoiety; whereby each of said aliphatic and heteroaliphatic moieties isindependently substituted or unsubstituted, branched or unbranched, orcyclic or acyclic, and each of said aryl, heteroaryl and heterocyclicmoieties is independently substituted or unsubstituted;
 108. The methodof claim 76, wherein any one or more of R₁, R_(A), R_(B), R_(C), orR_(D) is independently any one of the following groups:

wherein each occurrence of R_(J) is independently hydrogen, a protectinggroup, —OR_(K), ═O, —C(═O)R_(K), —CO₂R_(K), —CN, —SCN, halogen, —SR_(K),—SOR_(K), —SO₂R_(K), —NO₂, —N(R_(K))₂, —NHC(O)R_(K), —B(OR_(K))₂, or analiphatic, heteroaliphatic, aryl, or heteroaryl moiety, wherein eachoccurrence of R_(K) is independently hydrogen, or an aliphatic,heteroaliphatic, aryl, or heteroaryl moiety, or wherein two occurrencesof R_(K), taken together form a cyclic aliphatic or heteroaliphaticmoiety; wherein each occurrence of Y is independently O, S or NH;wherein each occurrence of x is independently 0-5; and wherein eachoccurrence of n is independently 0-3, or wherein R_(J) is a labelingreagent, whereby each of said aliphatic and heteroaliphatic moieties areindependently substituted or unsubstituted, branched or unbranched orcyclic or acyclic, and each of said aryl and heteroaryl moieties isindependently substituted or unsubstituted.
 109. The method of claim108, wherein R₁ is NR_(A)R_(B), R_(A) is hydrogen, R_(B) is —(C═O)R_(C),and R_(C) is iii, iv, vii, viii, ix, x, xv, or xvii, or R_(A) and R_(C)taken together form the structure of i or ii.
 110. The method of claim108, wherein R₁ is NR_(A)R_(B) and R_(A) is hydrogen, R_(B) is—(C═O)R_(C), and R_(C) is


111. The method of claim 108, 109, or 110, wherein R_(J) is hydrogen,halogen, —OH, lower alkyl or lower alkoxy.
 112. The method of claim 108,109, or 110, wherein R_(J) is a linker-biotin or a linker-fluoresceinmoiety.
 113. The method of claim 108, 109, or 110, wherein x is 1 or 2.114. The method of claim 76, wherein the compound of formula (I) has thestructure of any one of the formulae: